2-anilino-4-aminoalkyleneaminopyrimidines

ABSTRACT

The present invention relates to 2-arylamino-4-(aminoalkylene)aminopyrimidines inhibitors which are inhibitors and therefore inhibit Protein Kinase C-alpha (PKC-α). The PKC-α inhibitors of the present invention are important for improving myocardial intracellular calcium cycling, resulting in improved myocardial contraction and relaxation performance and thereby slowing the progression of heart failure. The present invention further relates to compositions comprising said 2-arylamino-4-(aminoalkylene)amino-pyrimidines and to methods for controlling, abating, or otherwise slowing the progression of heart failure.

RELATED CASES

This application is a continuation of U.S. application Ser. No. 11/762,406 filed Jun. 13, 2007, which claims priority benefit to U.S. provisional Application No. 60/813,875 filed Jun. 15, 2006.

FIELD OF THE INVENTION

The present invention relates to 2-arylamino-4-(aminoalkylene)aminopyrimidines which are inhibitors of Protein Kinase C-alpha (PKC-α). The PKC-α inhibitors of the present invention are important for improving myocardial intracellular calcium cycling, resulting in improved myocardial contraction and relaxation performance and thereby slowing the progression of heart failure. The present invention further relates to compositions comprising said 2-arylamino-4-(aminoalkylene)amino-pyrimidines and to methods for controlling, abating, or otherwise slowing the progression of heart failure.

BACKGROUND OF THE INVENTION

Many biologically active substances, for example, hormones, neurotransmitters and peptides are known to exert functions via intracellular mediators such as, cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), diacylglycerol (DAG) and calcium. In many cases, these mediators activate or inactivate intracellular kinases or phosphatases that are important in protein phosphorylation/dephosphorylation, and thus play important roles in regulating cellular processes and functions. The protein kinase C (PKC) family of calcium and/or lipid-activated serine-threonine kinases function downstream of nearly all membrane-associated signal transduction pathways.¹ Approximately 12 different isozymes comprise the PKC family, which are broadly classified by their activation characteristics. The conventional PKC isozymes (PKCα, βI, βII, and γ) are calcium- and lipid-activated, while the novel isozymes (ε, θ, η, and δ) and atypical isozymes (ζ, ι, ν, and μ) are calcium independent but activated by distinct lipids.² For example, stimulation of Gαq-coupled G-protein coupled receptors (GPCR) can activate phospholipase C (PLC) which in turn mediates hydrolysis of inositol phospholipids resulting in the generation of inositol 1,4,5-triphosphate (IP₃) and DAG. IP₃ and DAG can activate the different isoforms of PKC by mobilizing calcium (calcium sensitive enzymes) or by directly activating PKC, respectively. Once activated, PKC isozymes translocate to discrete subcellular locations through direct interactions with docking proteins termed RACKs (Receptor for Activated C Kinases), which permit specific substrate recognition and subsequent signal transduction.³

Alterations in PKC activity has been suggested to contribute to human diseases, inter alia, diabetes, numerous forms of cancer, microalbinuria, endothelial dysfunction, cerebrovascular disease, stroke, coronary heart disease, cardiovascular disease and sequela (e.g. arrhythmia, sudden death, increased infarct size, congestive heart failure, angina), myocardial ischemic states, hypertension, lipid disorders, ischemia-reperfusion injury, atherosclerosis, peripheral artery/vascular disease, microvascular complications of diabetes (neuropathy, nephropathy, retinopathy), restenosis, renal disease, blood coagulation disorders, inflammatory diseases and heart failure and inhibition of PKC in these settings could be used to treat or prevent human disease. Lending support to the modulation of PKC in cardiac disease, PKC activation has been associated with cardiac hypertrophy, dilated cardiomyopathy, ischemic injury and mitogen stimulation.

Heart disease is the leading cause of death in industrialized nations. Historically heart failure (HF) has been a product of hypertension, coronary heart disease, genetic disorders, valvular deformities, diabetes or cardiomyopathy. While the root cause of heart failure is multifaceted, it uniformly is marked by impaired diastolic and/or systolic function and can be accompanied by chamber enlargement which ultimately manifest in symptomatic heart failure (fatigue, pulmonary edema, circulatory congestion, etc.)

The risk of death due to heart failure is 5-10% annually in patients with mild symptoms of heart failure, and increases to 30-40% annually in patients with advanced heart failure, with a 50% overall mortality rate at 5 years. The current mainstays of heart failure therapy are drugs that act on the renin-angiotensin-aldosterone system (ACEI, ARB, aldosterone inhibitor), diuretics, digoxin and β-adrenergic receptor blockers. Despite the fact that multiple drug classes are used to treat heart failure patients, new cases of heart failure are growing at over 10% per year.

Patients with acute decompensated heart failure (ADHF) are a treatment challenge to physicians and can present with volume overload and/or diminished cardiac output. Initial treatments for ADHF patients include intravenous diuretics, vasodilators, natriuretic peptides and inotropic agents. Despite the widespread use of these agents, long-term safety and benefit of these drugs have been questioned. In the case of inotropes, drugs that increases cardiac output and cardiac contractility without increasing myocardial oxygen consumption or heart rate are desirous. Despite the available treatments for patients with ADHF, hospital readmission rates are approximately 50% within 6 months and mortality is approximately 20-40% at 1 year.

The primary function of the heart is to generate and sustain an arterial blood pressure necessary to provide adequate perfusion of organs. It has, therefore, become an area of intense investigation to decipher the mechanism(s) which initiate and contribute to the development of heart failure rather than relying on a means for treating the symptoms of heart failure alone. At the cardiomyocyte (cardiac contractile cells) level, impaired calcium cycling is a hallmark of heart failure as is the basis of contractile abnormalities. Calcium plays a key role in regulating kinases, phosphatases and transcription factors believed to influence the remodeling process indicating that both acute and sustained alterations in intracellular calcium levels may have profound effect on cardiac function and remodeling (i.e., changes in wall thickness or chamber volume). This theory would support the proposition that the development of new therapies addressing the slowing and preventing of the disease progression, would be perhaps more effective against heart failure than palliation of heart failure.

Therefore, there is a limited means to treat patients with various forms and stages of heart failure and there is incentive to develop novel, safe and effective treatments to prevent or treat patients with symptoms of heart failure, acute exacerbation of heart failure and chronic heart failure and other cardiovascular diseases. An agent that has benefits in the treating acute exacerbations of heart failure as well as treating chronic heart failure is desirous.

-   1. Molkentin et al. (2001) Annu. Rev. Physiol. 63:391-426. -   2. Dempsey et al. (2000) Am. J. Physiol. Lung Mol. Physiol.     279:247-251. -   3. Mochly-Rosen, D. (1995) Science 268:247-251.

SUMMARY OF THE INVENTION

The present invention meets the aforementioned needs in that it has been found that certain 2-arylamino-4-(aminoalkylene)aminopyrimidines are effective for inhibiting Protein Kinase C-alpha (PKC-α) thereby improving myocardial contraction and relaxation performance and slowing the progression of heart failure.

The present invention encompasses three major aspects each of which have their own separate categories, aspects, iterations, and specific iterative examples. The major aspects of the present invention include:

-   -   i) novel compositions of matter which are effective in         inhibiting PKC-α;     -   ii) compositions or pharmaceutical compositions (matrices)         comprising said compositions of matter;     -   iii) methods for controlling, abating, or alleviating one or         more of the causes of progressive heart failure which is         affected by administration of a PKC-α inhibitors, whether         administered alone or in a composition or within a         pharmaceutical composition (matrix); and     -   iv) a process for preparing the PKC-α inhibitors of the present         invention.

The first aspect of the present invention as a whole, relates to compounds, which include all enantiomeric and diastereomeric forms and pharmaceutically acceptable salts thereof, said compounds having the formula:

wherein R is a unit having the formula:

R² and R³ are each independently chosen from:

i) hydrogen; or

ii) C₁-C₄ substituted or unsubstituted linear, branched, or cyclic alkyl;

L is a linking unit having the formula:

—[C(R^(4a)R^(4b))]_(n)—

each R⁴ is independently chosen from

i) hydrogen; or

ii) C₁-C₄ alkyl;

the index n is from 1 to 4; and

R¹ is substituted or unsubstituted aryl.

The second major aspect of the present invention relates to compositions comprising:

-   -   a) an effective amount of one or more compounds according to the         present invention; and     -   b) one or more acceptable excipients.

The third major aspect of the present invention relates to methods of use. As described herein below, the PKC-α inhibitors of the present invention are important for improving myocardial contraction and relaxation performance and thereby slowing the progression of heart failure and their administration to humans is, therefore, an effective treatment for humans suffering from acute heart failure.

These and other objects, features, and advantages will become apparent to those of ordinary skill in the art from a reading of the following detailed description and the appended claims. All percentages, ratios and proportions herein are by weight, unless otherwise specified. All temperatures are in degrees Celsius (° C.) unless otherwise specified. All documents cited are in relevant part, incorporated herein by reference; the citation of any document is not to be construed as an admission that it is prior art with respect to the present invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention addresses several unmet medical needs, inter alia:

-   -   1) improving cardiac contraction/relaxation parameters in heart         failure patients, leading to reduction of symptoms; and     -   2) attenuating adverse cardiac remodeling in heart failure         patients, ultimately providing prolonged patient survival.

These and other unmet medical needs are resolved by the PKC-α inhibitors of the present invention, which are capable of blocking Protein Kinase C-alpha from impairing sarcoplasmic reticulum Ca²⁺ uptake. By providing heart failure patients with a PKC-α inhibitor, it is believed the patients will derive an improvement in cardiac function, thus resulting in improved myocardial contraction and relaxation performance and could result in slowing the progression to heart failure.

The following chemical hierarchy is used throughout the specification to describe and enable the scope of the present invention and to particularly point out and distinctly claim the units which comprise the compounds of the present invention. The term “hydrocarbyl” stands for any carbon atom-based unit (organic molecule), said units optionally containing one or more organic functional group, including inorganic atom comprising salts, inter alia, carboxylate salts, quaternary ammonium salts. Within the broad meaning of the term “hydrocarbyl” are the classes “acyclic hydrocarbyl” and “cyclic hydrocarbyl” which terms are used to divide hydrocarbyl units into cyclic and non-cyclic classes.

As it relates to the following definitions, “cyclic hydrocarbyl” units may comprise only carbon atoms in the ring (hydrocarbyl and aryl rings) or may comprise one or more heteroatoms in the ring (heterocyclic and heteroaryl). For “hydrocarbyl” rings the lowest number of carbon atoms in a ring are 3 carbon atoms; cyclopropyl. For “aryl” rings the lowest number of carbon atoms in a ring are 6 carbon atoms; phenyl. For “heterocyclic” rings the lowest number of carbon atoms in a ring is 1 carbon atom; diazirinyl, epoxy. For “heteroaryl” rings the lowest number of carbon atoms in a ring is 1 carbon atom; 1,2,3,4-tetrazolyl. The following is a non-limiting description of the terms “acyclic hydrocarbyl” and “cyclic hydrocarbyl” as used herein.

A. Substituted and Unsubstituted C₁-C₂₀ Acyclic Hydrocarbyl:

-   -   For the purposes of the present invention the term “substituted         and unsubstituted C₁-C₂₀ acyclic hydrocarbyl” encompasses 3         categories of units:

-   1) C₁-C₂₀ linear or branched alkyl, non-limiting examples of which     include, methyl (C₁), ethyl (C₂), n-propyl (C₃), iso-propyl (C₃),     n-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), tert-butyl (C₄), and     the like; substituted C₁-C₂₀ linear or branched alkyl, non-limiting     examples of which includes, hydroxymethyl (C₁), chloromethyl (C₁),     trifluoromethyl (C₁), aminomethyl (C₁), 1-chloroethyl (C₂),     2-hydroxyethyl (C₂), 1,2-difluoroethyl (C₂), 3-carboxypropyl (C₃),     and the like.

-   2) C₂-C₂₀ linear or branched alkenyl, non-limiting examples of which     include, ethenyl (C₂), 3-propenyl (C₃), 1-propenyl (also     2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),     buten-4-yl (C₄), and the like; substituted C₂-C₂₀ linear or branched     alkenyl, non-limiting examples of which include, 2-chloroethenyl     (also 2-chlorovinyl) (C₂), 4-hydroxybuten-1-yl (C₄),     7-hydroxy-7-methyloct-4-en-2-yl (C₉),     7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like.

-   3) C₂-C₂₀ linear or branched alkynyl, non-limiting examples of which     include, ethynyl (C₂), prop-2-ynyl (also propargyl) (C₃),     propyn-1-yl (C₃), and 2-methyl-hex-4-yn-1-yl (C₇); substituted     C₂-C₂₀ linear or branched alkynyl, non-limiting examples of which     include, 5-hydroxy-5-methylhex-3-ynyl (C₇),     6-hydroxy-6-methylhept-3-yn-2-yl (C₈), 5-hydroxy-5-ethylhept-3-ynyl     (C₉), and the like.

B. Substituted and Unsubstituted C₁-C₂₀ Cyclic Hydrocarbyl:

-   -   For the purposes of the present invention the term “substituted         and unsubstituted C₁-C₂₀ cyclic hydrocarbyl” encompasses 5         categories of units:

-   1) The term “carbocyclic” is defined herein as “encompassing rings     comprising from 3 to 20 carbon atoms, wherein the atoms which     comprise said rings are limited to carbon atoms, and further each     ring can be independently substituted with one or more moieties     capable of replacing one or more hydrogen atoms.” The following are     non-limiting examples of “substituted and unsubstituted C₃-C₂₀     carbocyclic rings” which encompass the following categories of     units:     -   i) carbocyclic rings having a single substituted or         unsubstituted hydrocarbon ring, non-limiting examples of which         include, cyclopropyl (C₃), 2-methyl-cyclopropyl (C₃),         cyclopropenyl (C₃), cyclobutyl (C₄), 2,3-dihydroxycyclobutyl         (C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),         cyclopentadienyl (C₅), cyclohexyl (C₆), cyclohexenyl (C₆),         cycloheptyl (C₇), cyclooctanyl (C₈), decalinyl (C₁₀),         2,5-dimethylcyclopentyl (C₅), 3,5-dichlorocyclohexyl (C₆),         4-hydroxycyclohexyl (C₆), and 3,3,5-trimethylcyclohex-1-yl (C₆).     -   ii) carbocyclic rings having two or more substituted or         unsubstituted fused hydrocarbon rings, non-limiting examples of         which include, octahydropentalenyl (C₈), octahydro-1H-indenyl         (C₉), 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl (C₉),         decahydroazulenyl (C₁₀); bicyclo[6.2.0]decanyl (C₁₀),         decahydronaphthalenyl (C₁₀), and dodecahydro-1H-fluorenyl (C₁₃).     -   iii) carbocyclic rings which are substituted or unsubstituted         bicyclic hydrocarbon rings, non-limiting examples of which         include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl,         bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl,         bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

-   2) The term “aryl” is defined herein as “units encompassing at least     one phenyl or naphthyl ring and wherein there are no heteroaryl or     heterocyclic rings fused to the phenyl or naphthyl ring and further     each ring can be independently substituted with one or more moieties     capable of replacing one or more hydrogen atoms.” The following are     non-limiting examples of “substituted and unsubstituted C₆-C₁₄ aryl     rings” which encompass the following categories of units:     -   i) C₆ or C₁₀ substituted or unsubstituted aryl rings; phenyl and         naphthyl rings whether substituted or unsubstituted,         non-limiting examples of which include, phenyl (C₆),         naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl         (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),         2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),         2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆),         3-methoxyphenyl (C₆), 8-hydroxynaphthylen-2-yl (C₁₀),         4,5-dimethoxynaphthylen-1-yl (C₁₀), and 6-cyano-naphthylen-1-yl         (C₁₀).     -   ii) C₆ or C₁₀ aryl rings fused with 1 or 2 saturated rings         non-limiting examples of which include,         bicyclo[4.2.0]octa-1,3,5-trienyl (C₈), and indanyl (C₉).

-   3) The terms “heterocyclic” and/or “heterocycle” are defined herein     as “units comprising one or more C₁-C₂₀ rings having from 3 to 20     atoms wherein at least one atom in at least one ring is a heteroatom     chosen from nitrogen (N), oxygen (O), or sulfur (S), or mixtures of     N, O, and S, and wherein further the ring which comprises the     heteroatom is also not an aromatic ring.” The following are     non-limiting examples of “substituted and unsubstituted C₁-C₂₀     heterocyclic rings” which encompass the following categories of     units:     -   i) heterocyclic units having a single ring containing one or         more heteroatoms, non-limiting examples of which include,         diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl         (C₃), pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl         (C₃), isoxazolinyl (C₃), isoxazolyl (C₃), thiazolidinyl (C₃),         isothiazolyl (C₃), isothiazolinyl (C₃), oxathiazolidinonyl (C₃),         oxazolidinonyl (C₃), hydantoinyl (C₃), tetrahydrofuranyl (C₄),         pyrrolidinyl (C₄), morpholinyl (C₄), piperazinyl (C₄),         piperidinyl (C₄), dihydropyranyl (C₅), tetrahydropyranyl (C₅),         piperidin-2-onyl (valerolactam) (C₅),         2,3,4,5-tetrahydro-1H-azepinyl (C₆), 2,3-dihydro-1H-indole (C₈),         and 1,2,3,4-tetrahydro-quinoline (C₉).     -   ii) heterocyclic units having 2 or more rings one of which is a         heterocyclic ring, non-limiting examples of which include         hexahydro-1H-pyrrolizinyl (C₇),         3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl (C₇),         3a,4,5,6,7,7a-hexahydro-1H-indolyl (C₈),         1,2,3,4-tetrahydroquinolinyl (C₉), and         decahydro-1H-cycloocta[b]pyrrolyl (C₁₀).

-   4) The term “heteroaryl” is defined herein as “encompassing one or     more C₁-C₂₀ rings comprising from 5 to 20 atoms wherein at least one     atom in at least one ring is a heteroatom chosen from nitrogen (N),     oxygen (O), or sulfur (S), or mixtures of N, O, and S, and wherein     further at least one of the rings which comprises a heteroatom is an     aromatic ring.” The following are non-limiting examples of     “substituted and unsubstituted C₁-C₂₀ heterocyclic rings” which     encompass the following categories of units:     -   i) heteroaryl rings containing a single ring, non-limiting         examples of which include, 1,2,3,4-tetrazolyl (C₁),         [1,2,3]triazolyl (C₂), [1,2,4]triazolyl (C₂), triazinyl (C₃),         thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl (C₃), furanyl (C₄),         thiopheneyl (C₄), pyrimidinyl (C₄), 2-phenylpyrimidinyl (C₄),         pyridinyl (C₅), 3-methylpyridinyl (C₅), and         4-dimethylaminopyridinyl (C₅).     -   ii) heteroaryl rings containing 2 or more fused rings one of         which is a heteroaryl ring, non-limiting examples of which         include: 7H-purinyl (C₅), 9H-purinyl (C₅), 6-amino-9H-purinyl         (C₅), 5H-pyrrolo[3,2-d]pyrimidinyl (C₆),         7H-pyrrolo[2,3-]pyrimidinyl (C₆), pyrido[2,3-]pyrimidinyl (C₇),         2-phenylbenzo[d]thiazolyl (C₇), 1H-indolyl (C₈),         4,5,6,7-tetrahydro-1-H-indolyl (C₈), quinoxalinyl (C₈),         5-methylquinoxalinyl (C₈), quinazolinyl (C₈), quinolinyl (C₉),         8-hydroxy-quinolinyl (C₉), and isoquinolinyl (C₉).

-   5) C₁-C₆ tethered cyclic hydrocarbyl units (whether C₃-C₁₀     carbocyclic units, C₆ or C₁₀ aryl units, C₁-C₁₀ heterocyclic units,     or C₁-C₁₀ heteroaryl units) which connected to another moiety, unit,     or core of the molecule by way of a C₁-C₆ alkylene unit.     Non-limiting examples of tethered cyclic hydrocarbyl units include     benzyl C₁-(C₆) having the formula:

-   -   wherein R^(a) is optionally one or more independently chosen         substitutions for hydrogen. Further examples include other aryl         units, inter alia, (2-hydroxyphenyl)hexyl C₆—(C₆);         naphthalen-2-ylmethyl C₁-(C₁₀), 4-fluorobenzyl C₁-(C₆),         2-(3-hydroxy-phenyl)ethyl C₂—(C₆), as well as substituted and         unsubstituted C₃-C₁₀ alkylenecarbocyclic units, for example,         cyclopropylmethyl C₁-(C₃), cyclopentylethyl C₂—(C₅),         cyclohexylmethyl C₁-(C₆). Included within this category are         substituted and unsubstituted C₁-C₁₀ alkylene-heteroaryl units,         for example a 2-picolyl C₁-(C₆) unit having the formula:

-   -   wherein R^(a) is the same as defined above. In addition, C₁-C₁₂         tethered cyclic hydrocarbyl units include C₁-C₁₀         alkyleneheterocyclic units and alkylene-heteroaryl units,         non-limiting examples of which include, aziridinylmethyl C₁-(C₂)         and oxazol-2-ylmethyl C₁-(C₃).

For the purposed of the present invention fused ring units, as well as spirocyclic rings, bicyclic rings and the like, which comprise a single heteroatom will be considered to belong to the cyclic family corresponding to the heteroatom containing ring. For example, 1,2,3,4-tetrahydroquinoline having the formula:

is, for the purposes of the present invention, considered a heterocyclic unit. 6,7-Dihydro-5H-cyclopentapyrimidine having the formula:

is, for the purposes of the present invention, considered a heteroaryl unit. When a fused ring unit contains heteroatoms in both a saturated and an aryl ring, the aryl ring will predominate and determine the type of category to which the ring is assigned. For example, 1,2,3,4-tetrahydro-[1,8]naphthyridine having the formula:

is, for the purposes of the present invention, considered a heteroaryl unit.

The term “substituted” is used throughout the specification. The term “substituted” is defined herein as “a hydrocarbyl moiety, whether acyclic or cyclic, which has one or more hydrogen atoms replaced by a substituent or several substituents as defined herein below.” The units, when substituting for hydrogen atoms are capable of replacing one hydrogen atom, two hydrogen atoms, or three hydrogen atoms of a hydrocarbyl moiety at a time. In addition, these substituents can replace two hydrogen atoms on two adjacent carbons to form said substituent, new moiety, or unit. For example, a substituted unit that requires a single hydrogen atom replacement includes halogen, hydroxyl, and the like. A two hydrogen atom replacement includes carbonyl, oximino, and the like. A two hydrogen atom replacement from adjacent carbon atoms includes epoxy, and the like. Three hydrogen replacement includes cyano, and the like. The term substituted is used throughout the present specification to indicate that a hydrocarbyl moiety, inter alia, aromatic ring, alkyl chain; can have one or more of the hydrogen atoms replaced by a substituent. When a moiety is described as “substituted” any number of the hydrogen atoms may be replaced. For example, 4-hydroxyphenyl is a “substituted aromatic carbocyclic ring”, (N,N-dimethyl-5-amino)octanyl is a “substituted C₈ alkyl unit, 3-guanidinopropyl is a “substituted C₃ alkyl unit,” and 2-carboxypyridinyl is a “substituted heteroaryl unit.”

The following are non-limiting examples of categories and examples herewith of units which can suitably substitute for hydrogen atoms on a cyclic or acyclic hydrocarbyl unit, described herein below as R⁵ units, wherein in the non-limiting examples provided herein below, R⁶ is hydrogen, C₁-C₁₀ linear or branched alkyl, C₂-C₁₀ linear or branched alkenyl, C₂-C₁₀ linear or branched alkynyl, and C₆ or C₁₀ aryl.

R⁵ units according to the present invention include the following:

-   -   i) —NHCOR⁶; for example, —NHCOCH₃, —NHCOCH₂CH₃, —NHCOC₆H₅;     -   ii) —COR⁶; for example, —COCH₃, —COCH₂CH₃, —COCH₂CH₂CH₃;     -   iii) —CO₂R⁶; for example, —CO₂CH₃, —CO₂CH₂CH₃, —CO₂CH₂CH₂CH₃;     -   iv) —OCOR⁶; for example, —OCOCH₃, —OCOCH₂CH₃, —OCOCH₂CH₂CH₃;     -   v) —C(═NH)NH₂;     -   vi) —NHC(═NH)NH₂;     -   vii) —N(R⁶)₂; for example, —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃);     -   viii) —NHC₆H₅;     -   ix) C₁-C₄ linear, branched, or cyclic alkyl; for example,         methyl, ethyl;     -   x) —CON(R⁶)₂; for example, —CONH₂, —CONHCH₃, —CON(CH₃)₂;     -   xi) —CONHNH₂;     -   xii) —NHCN;     -   xiii) —CN;     -   xiv) halogen: —F, —Cl, —Br, and —I;     -   xv) —NHN(R⁶)₂; for example, —NHNH₂, —NHNHCH₃, —NHN(CH₃)₂;     -   xvi) —OR⁶; for example, —OH, —OCH₃, —OCH₂CH₃, —OCH₂CH₂CH₃;     -   xvii) —NO₂;     -   xviii) —CH_(m)X_(n); wherein X is halogen, m is from 0 to 2,         m+n=3; for example, —CH₂F, —CHF₂, —CF₃, —CCl₃, or —CBr₃; and     -   xix) —SO₂N(R⁶)₂; for example, —SO₂NH₂; —SO₂NHCH₃; —SO₂NHC₆H₅.

For the purposes of the present invention the terms “compound” and “analog” stand equally well for the novel compositions of matter described herein, including all enantiomeric forms, diastereomeric forms, salts, and the like, and the terms “compound” and “analog” are used interchangeably throughout the present specification.

Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids. Other acids, such as oxalic acid, while not themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of this invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N—(C₁-C₄ alkyl)₄+salts.

The compounds of the present invention are 2-arylamino-4-(aminoalkylene)-aminopyrimidines having the core scaffold:

R units are amino units having the formula:

wherein R² and R³ are each independently chosen from:

-   -   i) hydrogen; or     -   ii) C₁-C₄ substituted or unsubstituted linear, branched, or         cyclic alkyl.

The first aspect of R units relates to amino units wherein R₂ and R₃ are independently chosen from hydrogen, methyl (C₁), or ethyl (C₂), said units having the formula:

-   -   i) —NH₂;     -   ii)-NHCH₃;     -   iii) —N(CH₃)₂;     -   iv) —NH(CH₂CH₃);     -   v) —N(CH₂CH₃)₂; and     -   vi) —N(CH₃)(CH₂CH₃).

The second aspect of R units relates to amino units wherein R² and R³ are independently chosen from hydrogen, n-propyl (C₃), or iso-propyl (C₃), said units having the formula:

-   -   i) —NH(CH₂CH₂CH₃);     -   ii) —N(CH₂CH₂CH₃)₂;     -   iii) —NH[CH(CH₃)₂];     -   iv) —N[CH(CH₃)₂]₂; and     -   v) —N(CH₂CH₂CH₃)[CH(CH₃)₂].     -   vi)

The third aspect of R units relates to amino units wherein R² is chosen from methyl (C₁) or ethyl (C₂), and R³ is chosen from n-propyl (C₃) or iso-propyl (C₃), said units having the formula:

-   -   i) —N(CH₃)(CH₂CH₂CH₃);     -   ii) —N(CH₂CH₃)(CH₂CH₂CH₃);     -   iii) —N(CH₃)[CH(CH₃)₂]; and     -   iv) —N(CH₂CH₃)[CH(CH₃)₂]₂.     -   v)     -   The fourth aspect of R units relates to amino units wherein R²         and R³ are independently chosen from hydrogen, n-butyl (C₄),         iso-butyl (C₄), sec-butyl (C₄), and tert-butyl (C₄),         non-limiting examples of said units having the formula:     -   i) —NH(CH₂CH₂CH₂CH₃);     -   ii) —N(CH₂CH₂CH₂CH₃)₂;     -   iii) —NH[CH₂CH(CH₃)₂];     -   iv) —N[CH₂CH(CH₃)₂]₂;     -   v) —NH[CH(CH₃)CH₂CH₃];     -   vi) —N[CH(CH₃)CH₂CH₃]₂;     -   vii) —NH[C(CH₃)₃];     -   viii) —N[C(CH₃)₃]₂     -   ix) —N(CH₂CH₂CH₂CH₃)[CH₂CH(CH₃)₂];     -   x) —N(CH₂CH₂CH₂CH₃)[CH(CH₃)CH₂CH₃]; and     -   xi) —N(CH₂CH₂CH₂CH₃)[C(CH₃)₃].

R¹ is substituted or unsubstituted aryl having the formula:

wherein R⁵ represents one or more (from 1 to 5) optionally present, and independently selected, substitutes for hydrogen as outlined herein above and described in the categories, aspects, iterations, examples, and tables herein below.

The first category of R¹ units relates to aryl units which are substituted by one or more R⁵ units chosen from:

-   -   i) C₁-C₄ linear, branched, or cyclic alkyl; for example, methyl         (C₁), ethyl (C₂), n-propyl (C₃), isopropyl (C₃), n-butyl (C₄),         and the like;     -   ii) halogen; for example, —F, —Cl, —Br, and —I;     -   iii) —OR⁶; for example, —OCH₃ (C₁), —OCH₂CH₃ (C₂), —OCH₂CH₂CH₃         (C₃), —OCH(CH₃)₂ (C₃), and the like;     -   iv) —SO₂N(R⁶)₂; for example, —SO₂NH₂, —SO₂NHCH₃, —SO₂NHC₆H₅, and         the like;     -   v) —CH_(m)X_(n); wherein each X is independently F, Cl, Br, or         I, m is from 0 to 2, m+n=3; for example, —CH₂F, —CHF₂, —CF₃,         —CCl₃, —CBr₃, and the like; and     -   vi) —NO₂;         wherein each R⁶ is independently hydrogen, or C₁-C₄ linear,         branched, or cyclic alkyl.

The first aspect of the first category of R¹ units relates to substituted phenyl units chosen from: 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 3-chloro-4-methyl-phenyl, 3-chloro-4-fluorophenyl, 3,4-difluorophenyl, 3-trifluoromethylphenyl, 3-trifluoro-methyl-4-chlorophenyl, 3-methoxyphenyl, 3-methylphenyl, 3-ethylphenyl, and 3-iso-propylphenyl.

The second aspect of the first category of R¹ units relates to substituted phenyl units chosen from: 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2-chlorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichloro-phenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl, 2,3,6-trichlorophenyl, 2,4,5-tri-chlorophenyl, and 2,4,6-trichlorophenyl.

The third aspect of the first category of R¹ units relates to substituted phenyl units chosen from: 2-methylphenyl, 4-methylphenyl, 2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethyl-phenyl, 2,6-dimethylphenyl, 3,4-dimethyl-phenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethyl-phenyl, 2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethyl-phenyl, 2-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl, 2,5-diethyl-phenyl, 2,6-diethylphenyl, 3,4-diethylphenyl, 2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl, 2,4,5-triethylphenyl, and 2,4,6-triethylphenyl.

The fourth aspect of the first category of R¹ units relates to substituted phenyl units chosen from: 2-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,4-dimethoxyphenyl, 2,3,4-trimethoxyphenyl, 2,3,5-trimethoxyphenyl, 2,3,6-trimethoxy-phenyl, 2,4,5-tri-methoxyphenyl, 2,4,6-trimethoxyphenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxy-phenyl, 2,3-dihydroxyphenyl, 2,4-dihydroxyphenyl, 2,5-dihydroxyphenyl, 2,6-dihydroxy-phenyl, 3,4-dihydroxy-phenyl, 2,3,4-trihydroxyphenyl, 2,3,5-trihydroxy-phenyl, 2,3,6-trihydroxyphenyl, 2,4,5-trihydroxyphenyl, and 2,4,6-trihydroxyphenyl.

However, other combinations of R⁵ units not specifically exemplified are encompassed within the first category of R¹.

The second category of R¹ units relates to aryl units which are substituted by one or more R⁵ units which are substituted or unsubstituted C₆ (phenyl) or C₁₀ (naphthyl) aryl units.

The first aspect of this category relates to R¹ units which are biphenyl wherein R⁵ is phenyl, for example, a compound having the formula:

The second aspect of this category relates to R¹ units which are substituted biphenyl non-limiting examples of which include 2′-fluoro-biphenyl-2-yl, 2′-fluoro-biphenyl-3-yl, 2′-fluoro-biphenyl-4-yl, 3′-fluoro-biphenyl-2-yl, 3′-fluoro-biphenyl-3-yl, 3′-fluoro-biphenyl-4-yl, 4′-fluoro-biphenyl-2-yl, 4′-fluoro-biphenyl-3-yl, 4′-fluoro-biphenyl-4-yl, 2′-chloro-biphenyl-2-yl, 2′-chloro-biphenyl-3-yl, 2′-chloro-biphenyl-4-yl, 3′-chloro-biphenyl-2-yl, 3′-chloro-biphenyl-3-yl, 3′-chloro-biphenyl-4-yl, 4′-chloro-biphenyl-2-yl, 4′-chloro-biphenyl-3-yl, 4′-chloro-biphenyl-4-yl, 2′-methyl-biphenyl-2-yl, 2′-methyl-biphenyl-3-yl, 2′-methyl-biphenyl-4-yl, 3′-methyl-biphenyl-2-yl, 3′-methyl-biphenyl-3-yl, 3′-methyl-biphenyl-4-yl, 4′-methyl-biphenyl-2-yl, 4′-methyl-biphenyl-3-yl, and 4′-methyl-biphenyl-4-yl.

However, other combinations of R⁵ units not specifically exemplified are encompassed within the second category of R¹.

The third category of R¹ units relates to phenyl units which are substituted by one or more R⁵ units which are substituted or unsubstituted C₃, C₄ or C₅ heteroaryl units.

The first aspect of this category relates to R¹ units which are C₄ or C₅ heteroaryl substituted phenyl units wherein R⁵ is, for example, pyridin-2-yl thereby forming a compound having the formula:

Non-limiting examples or this category of R¹ include 2-pyridin-2-yl-phenyl, 3-pyridin-2-yl-phenyl, 4-pyridin-2-yl-phenyl, 2-pyridin-3-yl-phenyl, 3-pyridin-3-yl-phenyl, 4-pyridin-3-yl-phenyl, 2-pyridin-4-yl-phenyl, 3-pyridin-4-yl-phenyl, 4-pyridin-4-yl-phenyl, 2-pyrimidin-2-yl-phenyl, 3-pyrimidin-2-yl-phenyl, 4-pyrimidin-2-yl-phenyl, 2-pyrimidin-3-yl-phenyl, 3-pyrimidin-3-yl-phenyl, 4-pyrimidin-3-yl-phenyl, 2-pyrimidin-4-yl-phenyl, 3-pyrimidin-4-yl-phenyl, and 4-pyrimidin-4-yl-phenyl.

The second aspect of this category relates to R¹ units which are C₃ heteroaryl substituted phenyl wherein R⁵ is, for example, 2-methyl-thiazol-4-yl thereby forming a compound having the formula:

Non-limiting examples of this aspect of the third category of R¹ include imidazol-1-yl, imidazol-2-yl, imidazol-4-yl, thiazol-2-yl, thiazol-4-yl, and the like.

L is a linking unit having the formula:

—[C(R^(4a)R^(4b))]_(n)—

wherein each R^(4a) and R^(4b) unit is independently chosen from:

-   -   i) hydrogen; or     -   ii) C₁-C₄ linear, branched, or cyclic alkyl; for example, methyl         (C₁), ethyl (C₂), n-propyl(C₃), iso-propyl (C₃), cyclopropyl         (C₃), n-butyl (C₄), iso-butyl (C₄), sec-butyl (C₄), and         tert-butyl (C₄);         the index n is from 1 to 4. The index n indicates the number of         units which comprise L linking units, for example, a linking         unit having the formula —CH₂— (methylene) would have an index n         equal to 1. A linking unit having the formula —CH₂CH₂—         (ethylene) or the unit having the formula —CH(CH₃)CH₂—         (2-propylene) each have an index n equal to 2.

The first category of L units relates to unsubstituted alkylene units chosen from:

-   -   i) —CH₂—, methylene;     -   ii) —CH₂CH₂—, ethylene;     -   iii) —CH₂CH₂CH₂—, propylene; and     -   iv) —CH₂CH₂CH₂CH₂—, butylene.

The first aspect of the first category of L units encompasses linking groups which are —CH₂CH₂CH₂—, propylene; as exemplified in the generic formula:

The second aspect of the first category of L units encompasses linking groups which are —CH₂CH₂—, ethylene.

The second category of L units relates to alkyl substituted alkylene units chosen from:

-   -   i) —CH(CH₃)CH₂—, 1-methylethylene;     -   ii) —CH₂CH(CH₃)—, 2-methylethylene;     -   iii) —CH(CH₃)CH₂CH₂—, 1-methylpropylene;     -   iv) —CH₂CH(CH₃)CH₂—, 2-methylpropylene; and     -   v) —CH₂C(CH₃)₂CH₂—, 2,2-dimethylpropylene.     -   The first aspect of the second category of L units encompasses         linking groups which are —CH(CH₃)CH₂CH₂—, 1-methylpropylene; as         exemplified in the generic formula:

wherein the above formula encompasses both the R and the S enantiomers of the linking unit.

Synthesis Procedure

The compounds of the present invention can be prepared by the following general procedure, the formulator adjusting the reaction conditions as is necessary and which one skilled in the art will be able to accomplish without undo experimentation.

Step 1: Preparation of intermediate 2-(methylthio)pyrimidine-4(3H)-one

This compound can be used in the preparation of each analog encompassed by the present invention. The general procedure follows.

To a solution of sodium hydroxide (8 g, 200 mmol) in H₂O (75 mL) at room temperature is added thiouridine (14.2 g, 100 mmol). The resulting mixture is stirred at room temperature for 20 minutes. Methyl iodide (6.86 mL, 110 mmol) is slowly added dropwise in THF (10 mL) and the mixture is stirred at room temperature for 18 hours. A white solid forms upon acidifying the mixture to pH 5 with glacial acetic acid. At this point the mixture is cooled in an ice bath and allowed to stand for approximately 2 hours after which the final product separates as a white solid and can be collected by filtration. First crop of crystals typically yields the desired product in an excess of 60% yield. ¹H NMR (DMSO-d₆, 300 MHz): δ 2.45 (s, 3H), 6.07 (d, J=6.6 Hz, 1H), 7.85 (d, J=6.6 Hz, 1H).

Step 2: Formation of 2-anilino Intermediate

2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one: To a 2-(methyl-thio)pyrimidine-4(3H)-one (14.2 g, 100 mmol) in diglyme (60 mL) is added the substituted or unsubstituted aniline of choice (200 mmol). The resulting mixture is heated to reflux and stirred for approximately 18 hours. The product which typically forms as a solid upon cooling the mixture to room temperature, is washed with solvent (pentane, hexane, or isopentane). However, solvent can be added to the reaction mixture to induce crystallization if necessary.

Step 3. Formation of 4-chloro-2-anilino Intermediate

2-(Substituted or unsubstituted phenylamino)-4-chloro-pyrimidine: To a 2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one (5.02 g, 22.6 mmol) and N,N-dimethyl-aniline (450 mL) is added of phosphorus oxychloride (450 mL). The resulting mixture is heated to reflux for 15 minutes, cooled to room temperature and concentrated in vacuo. The residue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with EtOAc (3×250 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. The residue can be conveniently purified over silica (5% EtOAc in hexanes) to afford the desired compound.

Alternatively, the 4-chloro-2-anilino intermediate can be synthesized the following way:

2-(Substituted or unsubstituted phenylamino)-4-chloro-pyrimidine: To a 2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one (3.00 g, 13.5 mmol) in toluene (30 mL) is added N,N-dimethyl-aniline (3.57 mL, 28.4 mmol) and phosphorus oxychloride (1.24 mL, 13.5 mmol). The resulting mixture is heated to reflux for 15 minutes, cooled to room temperature and neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with EtOAc (3×250 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. The residue can be conveniently purified over silica (5% EtOAc in hexanes) to afford the desired compound.

Step 4. Formation of Final Compounds (Analogs) of the Present Invention

To the 2-(Substituted or unsubstituted phenylamino)pyrimidin-4(3H)-one formed in Step (2) (100 mmol) in THF (500 mL) is added diisopropylethylamine (200 mmol) followed by the desired diamine (200 mmol). The resulting mixture is heated to reflux for approximately 18 hours. The reaction is cooled to room temperature and concentrated in vacuo. The residue which forms is diluted with water and extracted with solvent. The combined organic layers are dried (MgSO₄) and concentrated in vacuo. This residue can be crystallized or purified over silica to afford the final compound.

Schemes I-IV herein below provide illustrative examples of the preparation of compounds encompassed by the various categories of the present invention.

The analogs (compounds) of the present invention are arranged into several categories to assist the formulator in applying a rational synthetic strategy for the preparation of analogs which are not expressly exampled herein. The arrangement into categories does not imply increased or decreased efficacy for any of the compositions of matter described herein.

Analog Categories

The compounds which comprise Category I of the present invention are 2-(substituted phenylamino)-4-(amino- or substituted amino-propylene)aminopyrimidines having the formula:

wherein R, R¹, and R² are defined herein below in Table I.

TABLE I No R R¹ R² 1 3-chlorophenyl —H —H 2 3-chlorophenyl —CH₃ —H 3 3-chlorophenyl —CH₃ —CH₃ 4 3-chlorophenyl —CH₂CH₃ —H 5 3-chlorophenyl —CH₂CH₃ —CH₃ 6 3-chlorophenyl —CH₂CH₃ —CH₂CH₃ 7 4-chlorophenyl —H —H 8 4-chlorophenyl —CH₃ —H 9 4-chlorophenyl —CH₃ —CH₃ 10 4-chlorophenyl —CH₂CH₃ —H 11 4-chlorophenyl —CH₂CH₃ —CH₃ 12 4-chlorophenyl —CH₂CH₃ —CH₂CH₃ 13 3,4-dichlorophenyl —H —H 14 3,4-dichlorophenyl —CH₃ —H 15 3,4-dichlorophenyl —CH₃ —CH₃ 16 3,4-dichlorophenyl —CH₂CH₃ —H 17 3,4-dichlorophenyl —CH₂CH₃ —CH₃ 18 3,4-dichlorophenyl —CH₂CH₃ —CH₂CH₃ 19 3-chloro-4-methylphenyl —H —H 20 3-chloro-4-methylphenyl —CH₃ —H 21 3-chloro-4-methylphenyl —CH₃ —CH₃ 22 3-chloro-4-methylphenyl —CH₂CH₃ —H 23 3-chloro-4-methylphenyl —CH₂CH₃ —CH₃ 24 3-chloro-4-methylphenyl —CH₂CH₃ —CH₂CH₃ 25 3-chloro-2-fluorophenyl —CH₃ —CH₃ 26 3-chloro-2-fluorophenyl —CH₂CH₃ —CH₃ 27 3-chloro-2-fluorophenyl —CH₂CH₃ —CH₂CH₃ 28 3-chloro-4-fluorophenyl —CH₃ —CH₃ 29 3-chloro-4-fluorophenyl —CH₂CH₃ —CH₃ 30 3-chloro-4-fluorophenyl —CH₂CH₃ —CH₂CH₃ 31 3,4-difluorophenyl —H —H 32 3,4-difluorophenyl —CH₃ —H 33 3,4-difluorophenyl —CH₃ —CH₃ 34 3,4-difluorophenyl —CH₂CH₃ —H 35 3,4-difluorophenyl —CH₂CH₃ —CH₃ 36 3,4-difluorophenyl —CH₂CH₃ —CH₂CH₃ 37 3-trifluoromethylphenyl —H —H 38 3-trifluoromethylphenyl —CH₃ —H 39 3-trifluoromethylphenyl —CH₃ —CH₃ 40 3-trifluoromethylphenyl —CH₂CH₃ —H 41 3-trifluoromethylphenyl —CH₂CH₃ —CH₃ 42 3-trifluoromethylphenyl —CH₂CH₃ —CH₂CH₃ 43 3-trifluoromethyl-4-chlorophenyl —H —H 44 3-trifluoromethyl-4-chlorophenyl —CH₃ —H 45 3-trifluoromethyl-4-chlorophenyl —CH₃ —CH₃ 46 3-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —H 47 3-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —CH₃ 48 3-trifluoromethyl-4-chlorophenyl —CH₂CH₃ —CH₂CH₃ 49 3-methoxyphenyl —H —H 50 3-methoxyphenyl —CH₃ —H 51 3-methoxyphenyl —CH₃ —CH₃ 52 3-methoxyphenyl —CH₂CH₃ —H 53 3-methoxyphenyl —CH₂CH₃ —CH₃ 54 3-methoxyphenyl —CH₂CH₃ —CH₂CH₃ 55 3-methylphenyl —CH₃ —CH₃ 56 3-methylphenyl —CH₂CH₃ —CH₃ 57 3-methylphenyl —CH₂CH₃ —CH₂CH₃ 58 3-ethylphenyl —CH₃ —CH₃ 59 3-ethylphenyl —CH₂CH₃ —CH₃ 60 3-ethylphenyl —CH₂CH₃ —CH₂CH₃

The compounds which comprise Category I of the present invention can be prepared by the procedure outlined herein below in Scheme 1.

Example 1 N²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine (4)

Preparation of 2-(methylthio)pyrimidine-4(3H)-one (1): To a solution of sodium hydroxide (6.24 g, 156.07 mmol) in H₂O (55 mL) at room temperature is added thiouridine (10 g, 78.03 mmol). The resulting mixture is stirred at room temperature for 20 min. Methyl iodide (5.45 mL, 87.40 mmol) in THF (10 mL) is added dropwise slowly and the mixture is stirred at room temperature for 18 hours. A white solid forms upon acidifying the mixture to pH 5 with glacial acetic acid. The mixture is allowed to stand at 0° C. (ice bath) for 2 hours and filtered to afford 7.4 g (67% yield) of the desired compound as a white solid. ¹H NMR (DMSO-d₆, 300 MHz): δ 2.45 (s, 3H), 6.07 (d, J=6.6 Hz, 1H), 7.85 (d, J=6.6 Hz, 1H).

Preparation of 2-(3-chlorophenylamino)pyrimidin-4(3H)-one (2): To 2-(methyl-thio)pyrimidine-4(3H)-one, 1, (4.88 g, 34.37 mmol) in diglyme (20 mL) is added 3-chloroaniline (4.3 mL, 68.74 mmol). The resulting mixture is heated to reflux and stirred for 18 hours. A solid forms upon cooling the mixture to room temperature. The solid is washed with hexanes to afford 5.0 g (66% yield) of the desired compound. ¹H NMR (DMSO-d₆, 300 MHz): δ 5.91 (d, J=5.7 Hz, 2H), 7.05 (d, J=7.5 Hz, 1H), 7.11 (br s, 1H), 7.32 (t, J=7.8, 15.9 Hz, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.86 (d, J=4.5 Hz, 1H), 7.94 (s, 1H).

Preparation of 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine (3): To 2-(3-chlorophenylamino)pyrimidin-4(3H)-one, 2, (3.00 g, 13.5 mmol) in toluene (30 mL) is added N,N-dimethyl-aniline (3.57 mL, 28.4 mmol) and phosphorus oxychloride (1.24 mL, 13.5 mmol). The resulting mixture is heated to reflux for 15 minutes, cooled to room temperature and neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with EtOAc (3×250 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. The residue is purified over silica (5% EtOAc in hexanes) to afford 2.0 g (61% yield) of the desired compound. ¹H NMR (DMSO-d₆, 300 MHz): δ 7.06-7.04 (m, 2H), 7.34 (t, J=8.1, 1H), 7.65-7.61 (m, 1H), 7.93 (m, 1H), 8.50 (d, J=5.1 Hz, 1H), 10.26 (s, 1H).

Preparation of N²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine (4): To 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine, 3, (0.517 g, 2.16 mmol) in THF (10 mL) is added diisopropylethylamine (0.754 mL, 4.32 mmol) followed by 3-dimethyl-aminopropylamine (0.544 mL, 4.32 mmol). The resulting mixture is heated to reflux for 18 hours. The reaction is cooled to room temperature and concentrated in vacuo. The residue is diluted with 10 mL water and extracted with EtOAc (3×50 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. This residue is purified over silica (5% MeOH in CH₂Cl₂ with 0.7% Et₃N) to afford the desired compound as a yellow solid. The compound can be conveniently recrystallized from CH₂Cl₂ and MeOH to afford 0.206 g (40% yield) of a white solid. ¹H NMR (CD₃OD, 300 MHz): δ 1.85-1.95 (m, 2H). 2.40 (s, 6H), 2.59-2.65 (m, 2H), 3.40-3.51 (m, 2H), 5.99 (d, J=6.3 Hz, 1H), 6.95 (d, J=8.1 Hz, 1H), 7.24 (t, J=8.1 Hz, 1H), 7.40 (d, J=6.9 Hz, 1H), 7.78 (d, J=5.7 Hz, 1H), 8.03 (s, 1H). MS (ESI, pos. ion) m/z: 306 (M+1).

The following are non-limiting examples of compounds which comprise Category I of the present invention, the characterization of which will assist the formulator in establishing the chemical formulae of compounds which are not specifically exemplified herein.

N²-(3-chlorophenyl)-N⁴-(3-(diethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CD₃OD, 300 MHz): δ 1.30 (t, J=7.5 Hz, 6H), 2.15 (m, 2H), 3.18 (m, 6H), 3.60 (t, J=5.7 Hz, 1H), 6.15 (d, J=5.7 Hz, 1H), 6.98 (d, J=8.1 Hz, 1H), 7.23 (dd, J=8.1, 8.4 Hz, 1H), 7.40 (d, J=8.4 Hz, 1H), 7.81 (br d, J=5.7, 1H), 8.15 (s, 1H). MS (ESI, pos. ion) m/z: 334 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-methoxyphenyl)pyrimidine-2,4-diamine hydrochloride: ¹H NMR (CD₃OD, 300 MHz): δ 1.27 (t, J=7.5 Hz, 6H), 2.03-2.10 (m, 2H), 2.89 (s, 3H), 3.18-3.24 (m, 2H), 3.59-3.63 (m, 2H), 3.86 (s, 3H), 6.28 (d, J=7.5 Hz, 1H), 6.90 (d, J=8.4 Hz, 1H), 7.05 (d, J=8.1 Hz, 2H), 7.18 (s, 1H), 7.39 (t, J=7.8 Hz, 1H), 7.67 (d, J=7.2 Hz, 1H). MS (ESI, pos. ion) m/z: 302 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-methylphenyl)pyrimidine-2,4-diamine hydrochloride: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.98 (m, 2H), 2.33 (s, 3H), 2.72 (s, 6H), 3.09 (m, 2H), 3.45 (m, 2H), 6.24 (d, J=7.5 Hz, 1H), 6.97 (d, J=7.8 Hz, 1H), 7.29 (t, J=7.5 Hz, 1H), 7.40-7.43, (m, 4H), 7.84 (d, J=6.9 Hz, 1H), 9.17 (br sm 1H), 10.47 (br s, 2H). MS (ESI, pos. ion) m/z: 286 (M+1).

N⁴-(3-(dimethylamino)propyl)-N²-(3-(trifluoromethyl)phenyl)pyrimidine-2,4-diamine: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.63-1.72 (m, 2H), 2.13 (s, 6H), 2.26-2.31 (m, 2H), 3.32-3.34 (m, 2H), 5.98 (d, J=5.7 Hz, 1H), 7.17 (d, J=7.8 Hz, 1H), 7.30 (br s, 1H), 7.43 (t, J=7.8 Hz, 1H), 7.83 (br s, 1H), 8.48 (br s, 1H), 9.35 (br s, 1H). ¹⁹F 101.76 (s, 3F). MS (ESI, pos. ion) m/z: 340 (M+1).

N²-(3-chlorophenyl)-N⁴-(3-(methylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CD₃OD, 300 MHz): δ 2.02-2.11 (m, 2H), 2.72 (s, 3H), 3.06-3.11 (m, 2H), 3.60-3.65 (m, 2H), 6.32-6.34 (m, 1H), 7.29-7.51 (m, 3H), 7.72-7.79 (m, 2H). MS (ESI, pos. ion) m/z: 292 (M+1).

N⁴-(3-aminopropyl)-N²-(3-chlorophenyl)pyrimidine-2,4-diamine: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.90-1.97 (m, 2H), 2.89 (m, 2H), 3.46-3.52 (m, 2H), 6.35 (d, J=7.5 Hz, 1H), 7.22 (d, J=7.8 Hz, 1H), 7.42-7.53 (m, 2H), 7.88-7.92 (m, 2H), 8.12 (br s, 3H), 9.62 (br s, 1H), 11.15 (br s, 1H). MS (ESI, pos. ion) m/z: 278 (M+1).

N²-(3,4-difluorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.68 (m, 2H), 2.13 (s, 6H), 2.27 (m, 2H), 3.32 (m, 2H), 5.95 (d, J=5.4 Hz, 1H), 7.20-7.29 (m, 2H), 7.37-7.41 (m, 2H), 7.79 (m, 1H), 8.07-8.14 (m, 1H), 9.18 (s, 1H). ¹⁹F (DMSO-d₆, 300 MHz): δ 13.61 (m, 1F), 24.856 (m, 1F). MS (ESI, pos. ion) m/z: 308.27 (M+1).

N²-(3-chloro-4-methylphenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CD₃OD, 300 MHz): δ 1.85 (m, 2H), 2.28 (s, 6H), 2.32 (s, 3H), 2.48 (t, 2H), 3.46 (m, 2H), 5.96 (d, J=6.3 Hz, 1H), 7.16 (d, J=8.1 Hz, 1H), 7.29 (d, J=5.4 Hz, 1H), 7.75 (d, J=5.1 Hz, 1H), 8.00 (s, 1H). MS (ESI, pos. ion) m/z: 320 (M+1).

N²-(3,4-dichlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CDCl₃, 300 MHz): δ 1.85 (m, 2H), 2.34 (s, 6H), 2.52 (t, J=6.0 Hz, 2H), 3.50 (m, 2H), 5.91 (d, J=6.0 Hz, 1H), 6.21 (s, 1H), 7.10 (s, 1H), 7.30 (d, J=6.0 Hz, 1H), 7.92 (d, J=5.7 Hz, 1H), 8.13 (s, 1H). MS (ESI, pos. ion) m/z: 340 (M+1).

N²-(3-chloro-4-fluorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CDCl₃, 300 MHz): δ 1.79-1.86 (m, 2H), 2.30 (s, 6H), 2.46 (t, J=6.0 Hz, 2H), 3.46 (m, J=6.0 Hz, 2H), 5.89 (d, J=6.0 Hz, 1H), 6.13 (s, 1H), 7.06 (t, J=6.0 Hz, 1H), 7.17 (s, 1H), 7.27 (m, 1H), 7.26 (d, J=6.6 Hz, 1H), 7.29 (s, 1H), 7.92 (d, J=6.0 Hz, 1H), 8.02 (d, J=6.6 Hz, 1H). HRMS calcd for C₁₅H₁₉ClFN₅ 324.1391 m/z (M+1)⁺; observed 324.1399 m/z.

N²-(4-chloro-3-(trifluoromethyl)phenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine: ¹H NMR (CDCl₃, 300 MHz): δ 1.80 (m, 2H), 2.27 (s, 6H), 2.45 (t, J=6.0 Hz, 2H), 3.49 (bs, 2H), 5.91 (d, J=6.0 Hz, 1H), 6.37 (t, J=4.8 Hz, 1H)), 7.37 (d, J=8.7 Hz, 1H), 7.58 (d, J=8.7 Hz, 1H), 7.94 (d, J=6.0 Hz, 1H), 1H), 8.34 (bs, 2H). HRMS calcd for C₁₆H₁₉ClF₃N₅ 374.1359 m/z (M+H)⁺; observed 374.1357 m/z.

N⁴-(3-(dimethylamino)propyl)-N²-(3-ethylphenyl)pyrimidine-2,4-diamine: ¹H NMR (CD₃OD, 300 MHz): δ 1.29 (t, J=7.8 Hz, 3H), 2.07 (m, 2H), 2.71 (q, J=7.8 Hz, 2H)), 2.85 (s, 6H), 3.16 (m, 2H), 3.60 (t, J=6.6 Hz, 2H), 6.24 (d, J=7.5 Hz, 1H), 7.16 (m, 1H), 7.34 (s, 1H), 7.38 (m, 2H), 7.65 (d, J=7.2 Hz, 1H). HRMS calcd for C₁₇H₂₅N₅ 300.2188 m/z (M+1)⁺; observed 300.2194 m/z.

N²-(2-fluoro-3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine ¹H NMR (CDCl₃, 300 MHz): δ 1.75-1.82 (m, 2H), 2.28 (s, 6H), 2.44 (t, J=6.3 Hz, 2H), 3.46 (s, 2H), 5.92 (d, J=6.0 Hz, 1H), 6.02 (bs, 1H), 6.97-7.08, (m, 3H), 7.95 (d, J=5.1 Hz, 1H), 8.49 (t, J=7.8 Hz, 1H). HRMS calcd for C₁₅H₁₉N₅FCl, 324.1391 m/z (M+H)⁺; observed 324.1394 m/z.

Further compounds which are encompassed within Category I of the present invention but which are not fully exemplified include:

-   N⁴-(3-(dimethylamino)propyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(3-(dimethylamino)propyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine;     and -   N⁴-(3-(dimethylamino)propyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine.

The compounds which comprise Category II of the present invention are 2-(substituted phenylamino)-4-(amino- or substituted amino-ethylene, or substituted amino-butylene)-aminopyrimidines having the formula:

wherein R², R³, R⁵, and L are defined herein below in Table II.

TABLE II No. L R² R³ R⁵ 61 —CH₂— —H —H 3-chloro 62 —CH₂— —CH₃ —H 3-chloro 63 —CH₂— —CH₃ —CH₃ 3-chloro 64 —CH₂— —CH₂CH₃ —H 3-chloro 65 —CH₂— —CH₂CH₃ —CH₃ 3-chloro 66 —CH₂— —CH₂CH₃ —CH₂CH₃ 3-chloro 67 —CH₂CH₂— —H —H 3-chloro 68 —CH₂CH₂— —CH₃ —H 3-chloro 69 —CH₂CH₂— —CH₃ —CH₃ 3-chloro 70 —CH₂CH₂— —CH₂CH₃ —H 3-chloro 71 —CH₂CH₂— —CH₂CH₃ —CH₃ 3-chloro 72 —CH₂CH₂— —CH₂CH₃ —CH₂CH₃ 3-chloro 73 —CH₂C(CH₃)₂CH₂— —H —H 3-trifluoromethyl 74 —CH₂C(CH₃) ² CH ² — —CH₃ —H 3-trifluoromethyl 75 —CH₂C(CH₃) ² CH ² — —CH₃ —CH₃ 3-trifluoromethyl 76 —CH₂C(CH₃) ² CH ² — —CH₂CH₃ —H 3-trifluoromethyl 77 —CH₂C(CH₃) ² CH ² — —CH₂CH₃ —CH₃ 3-trifluoromethyl 78 —CH₂C(CH₃)₂CH₃₂— —CH₂CH₃ —CH₂CH₃ 3-trifluoromethyl 79 —CH₂CH₂CH₂CH₂— —H —H 3-chloro 80 —CH₂CH₂CH₂CH₂— —CH₃ —H 3-chloro 81 —CH₂CH₂CH₂CH₂— —CH₃ —CH₃ 3-chloro 82 —CH₂CH₂CH₂CH₂— —CH₂CH₃ —H 3-chloro 83 —CH₂CH₂CH₂CH₂— —CH₂CH₃ —CH₃ 3-chloro 84 —CH₂CH₂CH₂CH₂— —CH₂CH₃ —CH₂CH₃ 3-chloro

The compounds which comprise Category II of the present invention can be prepared by the procedure outlined herein below in Schemes II and III and Examples 3 and 4.

Example 2 N⁴-[3-(dimethylamino)-2,2-dimethylpropyl]-N²-[3-(trifluoromethyl)phenyl]pyrimidine-2,4-diamine Hydrochloride (7)

Preparation of 2-(3-trifluoromethylphenylamino)pyrimidin-4(3H)-one (5): To 2-(methyl-thio)pyrimidine-4(3H)-one, 1, (5 g, 35.21 mmol) in diglyme (25 mL) is added 3-trifluoromethylaniline (5.26 mL, 42.25 mmol). The resulting mixture is heated to reflux and stirred for 18 hours. A solid forms upon cooling the mixture to room temperature. The solid is washed with hexanes to afford 1.9 g (21% yield) of the desired compound. MS (ESI, pos. ion) m/z: 256 (M+1).

Preparation of 4-chloro-N-(3-trifluoromethylphenyl)pyrimidin-2-amine (6): To 2-(3-chloro-phenylamino)pyrimidin-4(3H)-one, 5, (1.9 g, 7.4 mmol) and N,N-dimethyl-aniline (2 mL) is added of phosphorus oxychloride (20 mL). The resulting mixture is heated to reflux for 15 minutes, cooled to room temperature and concentrated in vacuo. The residue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with ethyl acetate (3×50 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. The residue is purified over silica (5% EtOAc in hexanes) to afford 0.62 g (31% yield) of the desired compound. ¹H NMR (DMSO-d₆, 300 MHz): δ 7.07 (d, J=6.3 Hz, 1H), 7.34 (d, J=8.4 Hz, 1H), 7.55 (t, J=8.1 Hz, 1H), 7.96 (d, J=8.4 Hz, 1H), 8.20 (s, 1H), 8.52 (d, J=6.0 Hz, 1H), 10.40 (s, 1H). ¹⁹F NMR (DMSO-d₆, 282 MHz): δ 101.67 (s, 3F).

Preparation of N⁴-[3-(dimethylamino)-2,2-dimethylpropyl]-N²-[3-(trifluoro-methyl)phenyl]pyrimidine-2,4-diamine hydrochloride (7): 4-Chloro-N-(3-(trifluoro-methyl)phenyl)pyrimidin-2-amine, 6, (200 mg, 0.73 mmol), N¹,N¹,2,2-tetramethyl-propane-1,3-diamine (0.232 mL, 1.46 mmol), and diisopropylethylamine (0.254 mL, 1.46 mmol) are dissolved in THF (5 mL) and heated to reflux for 20 hours. The reaction is cooled to room temperature and partitioned between EtOAc and water. The organic layer is dried (MgSO₄), concentrated in vacuo, and purified over silica (5% MeOH ramped to 6% MeOH in CH₂Cl₂ with 0.7% added triethylamine) to give an oil. The oil is dissolved in MeOH (0.5 mL) and Et₂O (20 mL) and cooled to 0° C. To this solution is added a solution of 4M HCl in dioxane (0.25 mL) in one portion. The white precipitate which forms is collected by filtration and dried at reduced pressure to afford the desired compound as a white solid: ¹H NMR (DMSO-d₆, 300 MHz) δ 1.09 (s, 6H), 2.81 (s, 3H), 2.83 (s, 3H), 3.11 (d, J=4.2 Hz, 2H), 3.38-3.50 (m, 2H), 6.48 (d, J=7.2 Hz, 1H), 7.51 (d, J=7.5 Hz, 1H), 7.66 (t, J=7.5 Hz, 1H), 7.74 (d. J=6.6 Hz, 1H), 7.91 (d, J=7.2 Hz, 1H), 8.20 (s, 1H), 9.67 (bs, 1H), 10.80 (bs, 1H); HRMS calcd for C₁₈H₂₄F₃N₅, 368.2062 m/z (M+H)⁺; observed 368.2072 m/z.

Example 3 N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine (8)

Preparation of N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine(8): To 4-chloro-N-(3-chlorophenyl)pyrimidin-2-amine (0.2 g, 0.84 mmol) in THF (4 mL) is added diisopropylethylamine (0.29 mL, 1.67 mmol) followed by 4-dimethylaminobutylamine (0.2 g, 1.67 mmol). The resulting mixture is heated to reflux for 6 hours. Another 2 equivalents of 4-dimethylaminobutylamine (0.2 g, 1.67 mmol) are added and the reaction heated at reflux for 18 hours. The reaction is cooled to room temperature and concentrated in vacuo. The residue is diluted with water (5 mL) and extracted with EtOAc (3×25 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. This residue is purified over silica (5% MeOH in CH₂Cl₂ with 0.7% Et₃N) to afford 7 mg (3% yield) of the desired compound. ¹H NMR (CD₃OD, 300 MHz): δ 1.62-1.66 (m, 4H), 2.26 (s, 6H), 2.39-2.44 (m, 2H), 3.46-3.48 (m, 2H), 5.97 (d, J=5.7 Hz, 1H), 6.93 (d, J=8.1 Hz, 1H), 7.22 (t, J=8.1 Hz, 1H), 7.40 (d, J=8.1 Hz, 1H), 7.75 (d, J=5.7 Hz, 1H), 8.06 (s, 1H). MS (ESI, pos. ion) m/z: 320 (M+1).

The following are non-limiting examples of compounds which comprise Category II of the present invention, the characterization of which will assist the formulator in establishing the chemical formulae of compounds which are not specifically exemplified herein.

N²-(3-chlorophenyl)-N⁴-(2-(dimethylamino)ethyl)pyrimidine-2,4-diamine. ¹H NMR (CD₃OD, 300 MHz): δ 2.33 (s, 6H), 2.62 (t, J=6.0 Hz, 2H), 3.58 (t, J=6.0 Hz, 2H), 5.99 (d, J=6.0 Hz, 1H), 6.95 (d, J=8.1 Hz, 1H), 7.23 (t, J=8.1 Hz, 1H), 7.44 (d, J=8.1 Hz, 1H), 7.78 (d, J=6.0 Hz, 1H), 7.95 (s, 1H). MS (ESI, pos. ion) m/z: 292 (M+1).

N⁴-(3-aminopropyl)-N²-(3-chlorophenyl)-N⁴-methylpyrimidine-2,4-diamine: ¹H NMR (DMSO-d₆, 300 MHz): δ 1.88-1.92 (m, 2H), 2.78-2.84 (m, 2H), 3.13 (s, 3H), 3.62-3.68 (m, 2H), 3.43 (br s, 1H), 7.06-7.08 (m, 1H), 7.30-7.38 (m, 1H), 7.50-7.53 (m, 1H), 7.93-8.04 (m, 4H), 10.25 (br s, 1H). MS (ESI, pos. ion) m/z: 292 (M+1).

Further compounds which are encompassed within Category II of the present invention but which are not fully exemplified include:

-   N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(3-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(2-(dimethylamino)ethyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine; -   N²-(3-chlorophenyl)-N⁴-(4-(dimethylamino)butyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(3-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(4-fluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-difluorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2-chlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(4-chlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-dichlorophenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(4-methylphenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,3-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,4-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,5-dimethylphenyl)pyrimidine-2,4-diamine; -   N⁴-(4-(dimethylamino)butyl)-N²-(2,6-dimethylphenyl)pyrimidine-2,4-diamine;     and -   N⁴-(4-(dimethylamino)butyl)-N²-(3,4-dimethylphenyl)pyrimidine-2,4-diamine.

The compounds which comprise Category III of the present invention are 2-(substituted phenylamino)-4-(amino- or substituted amino-propylene)aminopyrimidines having the formula:

wherein R⁵ is substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, or substituted or unsubstituted heterocyclic, and R¹, and R² are defined herein below in Table IIII.

TABLE III No R⁵ R¹ R² 85 3-phenyl —H —H 86 3-phenyl —CH₃ —H 87 3-phenyl —CH₃ —CH₃ 88 3-phenyl —CH₂CH₃ —H 89 3-phenyl —CH₂CH₃ —CH₃ 90 3-phenyl —CH₂CH₃ —CH₂CH₃ 91 4-phenyl —H —H 92 4-phenyl —CH₃ —H 93 4-phenyl —CH₃ —CH₃ 94 4-phenyl —CH₂CH₃ —H 95 4-phenyl —CH₂CH₃ —CH₃ 96 4-phenyl —CH₂CH₃ —CH₂CH₃ 97 thiazol-4-yl —H —H 98 thiazol-4-yl —CH₃ —H 99 thiazol-4-yl —CH₃ —CH₃ 100 thiazol-4-yl —CH₂CH₃ —H 101 thiazol-4-yl —CH₂CH₃ —CH₃ 102 thiazol-4-yl —CH₂CH₃ —CH₂CH₃ 103 2-methylthiazol-4-yl —H —H 104 2-methylthiazol-4-yl —CH₃ —H 105 2-methylthiazol-4-yl —CH₃ —CH₃ 106 2-methylthiazol-4-yl —CH₂CH₃ —H 107 2-methylthiazol-4-yl —CH₂CH₃ —CH₃ 108 2-methylthiazol-4-yl —CH₂CH₃ —CH₂CH₃ 109 imidazol-2-yl —H —H 110 imidazol-2-yl —CH₃ —H 111 imidazol-2-yl —CH₃ —CH₃ 112 imidazol-2-yl —CH₂CH₃ —H 113 imidazol-2-yl —CH₂CH₃ —CH₃ 114 imidazol-2-yl —CH₂CH₃ —CH₂CH₃ 115 imidazol-4-yl —H —H 116 imidazol-4-yl —CH₃ —H 117 imidazol-4-yl —CH₃ —CH₃ 118 imidazol-4-yl —CH₂CH₃ —H 119 imidazol-4-yl —CH₂CH₃ —CH₃ 120 imidazol-4-yl —CH₂CH₃ —CH₂CH₃ 121 2-methylimidazol-4-yl —H —H 122 2-methylimidazol-4-yl —CH₃ —H 123 2-methylimidazol-4-yl —CH₃ —CH₃ 124 2-methylimidazol-4-yl —CH₂CH₃ —H 125 2-methylimidazol-4-yl —CH₂CH₃ —CH₃ 126 2-methylimidazol-4-yl —CH₂CH₃ —CH₂CH₃

The compounds which comprise Category III of the present invention can be prepared by the following procedures outlined herein below in Scheme IV and V and Examples 4 and 5.

Example 4 N⁴-(3-(dimethylamino)propyl)-N²-m-biphenylpyrimidine-2,4-diamine hydrochloride (11)

Preparation of 2-(3-biphenylamino)pyrimidin-4(3H)-one (9): To 2-(methylthio)-pyrimidine-4(3H)-one, 1, (790 mg, 5.5 mmol) in diglyme (5 mL) is added 3-amino-biphenyl (1.91 g, 11.2 mmol). The resulting mixture is stirred at reflux for 18 hours. The mixture is cooled to room temperature and hexane is added and a precipitate forms which is collected by filtration to afford 1.34 g (92% yield) of the desired compound which is used without further purification. MS (ESI, pos. ion) m/z: 264 (M+1).

Preparation of 4-chloro-N-(3-biphenyl)pyrimidin-2-amine (10): To 2-(3-biphenyl-amino)pyrimidin-4(3H)-one, 9, (1.34 g, 5.0 mmol), and N,N-dimethylaniline (1.5 mL) is added phosphorus oxychloride (10 mL). The resulting mixture is heated to reflux, stirred for 1 hour, cooled to room temperature and concentrated in vacuo. The residue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with EtOAc (2×50 mL). The combined organic layers are washed once with brine, dried (MgSO₄), and concentrated in vacuo. The residue is purified over silica (5% EtOAc in hexanes) to afford 780 mg (54% yield) of the desired compound. MS (ESI, pos. ion) m/z: 282 (M+1).

Preparation of N²-(3-biphenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine hydrochloride (11): To 4-chloro-N-(3-biphenyl)pyrimidin-2-amine, 10, (102 mg, 0.38 mmol) in THF (5 mL) is added N,N-diisopropylethylamine (0.15 mL, 0.84 mmol) followed by 3-(dimethylamino)propylamine (0.15 mL, 1.2 mmol). The resulting mixture is heated to reflux for 48 hours. The reaction is cooled to room temperature and concentrated in vacuo to afford a residue which is purified over silica (95:3:2 CH₂Cl₂/MeOH/7N NH₃ in MeOH) to afford 61 mg (48% yield) of the desired compound. The hydrochloride salt can be formed by treatment of the neutral compound with HCl in dioxane (4M). ¹H NMR (CD₃OD, 300 MHz): 6(ppm) 1.74 (m, 2H), 2.15 (s, 6H), 2.27 (t, J=6.0 Hz, 2H), 3.40 (m, 2H), 5.94 (d, J=6.0 Hz, 1H), 7.19 (d, J=7.2 Hz, 1H), 7.34 (m, 2H), 7.42 (m, 2H), 7.53 (d, J=7.5 Hz, 1H), 7.61 (m, 2H), 7.76 (d, J=5.4 Hz, 1H), 8.12 (s, 1H). MS (ESI, pos. ion) m/z: 348 (M+1).

Example 5 N⁴-(3-(dimethylamino)propyl)-N²-(3-(2-methylthiazol-4-yl)phenyl)pyrimidine-2,4-diamine Hydrochloride (13)

Preparation of 2-(3-(2-methylthiazol-4-yl)phenylamino)pyrimidin-4(3H)-one (11): To 2-(methyl-thio)pyrimidine-4(3H)-one, 1, (1 g, 14.08 mmol) in diglyme (9 mL) is added 3-(2-methylthiazol-4-yl)benzenamine (3.20 g, 16.90 mmol). The resulting mixture is heated to reflux and stirred for 18 hours. A solid forms upon cooling the mixture to room temperature. The solid is washed with hexanes to afford 1.0 g (25% yield) of the desired compound. MS (ESI, pos. ion) m/z: 285 (M+1).

Preparation of 4-chloro-N-(3-(2-methylthiazol-4-yl)phenyl)pyrimidin-2-amine (12): To 2-(3-(2-methylthiazol-4-yl)phenylamino)pyrimidin-4(3H)-one, 11, (1.0 g, 3.5 mmol) and N,N-dimethyl-aniline (1 mL) is added phosphorus oxychloride (9.5 mL). The resulting mixture is heated to reflux for 15 minutes, cooled to room temperature and concentrated in vacuo. The residue is neutralized to pH 7 with 1M NaOH (aqueous). The organic layer is extracted with ethyl acetate (3×50 mL). The combined organic layers are dried (MgSO₄) and concentrated in vacuo. The residue is purified over silica (5% EtOAc in hexanes) to afford 0.50 g (47% yield) of the desired compound. ¹H NMR (DMSO-d₆, 300 MHz): δ 2.73 (s, 3H), 6.98 (d, J=5.1 Hz, 1H), 7.37 (t, J=8.1 Hz, 1H), 7.57 (d, J=8.1 Hz, 1H), 7.72 (d, J=8.1 Hz, 1H), 7.83 (s, 1H), 8.25 (s, 1H), 8.46 (d, J=5.1 Hz, 1H), 10.08 (s, 1H). MS (ESI, pos. ion) m/z: 303 (M+1).

Preparation of N⁴-(3-(dimethylamino)propyl)-N²-(3-(2-methylthiazol-4-yl)phenyl)pyrimidine-2,4-diamine hydrochloride (13): 4-chloro-N-(3-(2-methylthiazol-4-yl)phenyl)pyrimidin-2-amine, 12, (400 mg, 1.30 mmol), N¹,N¹-dimethylpropane-1,3-diamine (0.25 mL, 2.0 mmol), and diisopropylethylamine (0.46 mL, 2.6 mmol) are dissolved in THF (10 mL) and heated to reflux for 20 hours. The reaction is cooled to room temperature and partitioned between EtOAc and water. The organic layer is dried (MgSO₄), concentrated in vacuo, and purified over silica (5% MeOH ramped to 6% MeOH in CH₂Cl₂ with 0.7% added triethylamine) to give an oil. The oil is dissolved in MeOH (0.5 mL) and Et₂O (20 mL) and cooled to 0° C. To this solution is added a solution of 4M HCl in dioxane (0.25 mL) in one portion. The white precipitate which forms is collected by filtration and dried at reduced pressure to afford the desired compound as a white solid: ¹H NMR (CD₃OD, 300 MHz) δ 2.03-2.08 (m, 2H), 2.79 (s, 6H), 2.80 (s, 3H), 3.10-3.15 (m, 2H), 3.65 (t, J=6.6 Hz, 2H), 6.30 (d, J=7.5 Hz, 1H), 7.45 (d, J=7.8 Hz, 1H), 7.54 (t, J=7.8 Hz, 1H), 7.70 (d. J=7.5 Hz, 1H), 7.77 (s, 1H), 7.82 (d, J=7.8 Hz, 1H), 8.19 (s, 1H); HRMS calcd for C₁₉H₂₄N₆S, 369.1861 m/z (M+H)⁺; observed 369.1855 m/z.

The compounds of the present invention are inhibitors of Protein Kinase C-alpha (PKC-α), therefore, they are PKC-α inhibitors which are capable of improving myocardial contraction and relaxation performance and slow the progression of heart failure. The compounds potentially inhibit additional isoforms of conventional PKC, such as PKC-β or PKc-γ. This is not undesired and can lead to increased pharmacological effects.

The level of disease, for example, the relative degree of heart failure due to PKC-α activity will vary from patient to patient and be predicated by other exacerbating circumstances, inter alia, presence of other disease conditions (diabetes, high blood pressure, and the like) or patients may suffer from other conditions such as obesity. Therefore, the formulator may be required to employ differing levels or amounts of the compounds described herein to obtain a therapeutic level. The formulator can determine this amount by any of the known testing procedures known to the artisan.

Formulations

The present invention also relates to compositions or formulations which comprise the PKC-α inhibitors according to the present invention. In general, the compositions of the present invention comprise:

-   -   a) an effective amount of one or more         2-arylamino-4-(aminoalkylene)amino-pyrimidines or salts thereof         according to the present invention which are effective for         inhibiting PKC-α; and     -   b) one or more excipients.     -   For the purposes of the present invention the term “excipient”         and “carrier” are used interchangeably throughout the         description of the present invention. One aspect of excipient         and carrier relate to their definition in terms of a medicament,         said terms are defined in that respect as, “ingredients which         are used in the practice of formulating a safe and effective         pharmaceutical composition.”

The formulator will understand that excipients are used primarily to serve in delivering a safe, stable, and functional pharmaceutical, serving not only as part of the overall vehicle for delivery but also as a means for achieving effective absorption by the recipient of the active ingredient. An excipient may fill a role as simple and direct as being an inert filler, or an excipient as used herein may be part of a pH stabilizing system or coating to insure delivery of the ingredients safely to the stomach. The formulator can also take advantage of the fact the compounds of the present invention have improved cellular potency, pharmacokinetic properties, as well as improved oral bioavailability.

Non-limiting examples of compositions according to the present invention include:

-   -   a) from about 0.001 mg to about 1000 mg of one or more PKC-α         inhibitors according to the present invention; and     -   b) one or more excipient.

Another embodiment according to the present invention relates to the following compositions:

-   -   a) from about 0.01 mg to about 100 mg of one or more PKC-α         inhibitors according to the present invention; and     -   b) one or more excipient.

A further embodiment according to the present invention relates to the following compositions:

-   -   a) from about 0.1 mg to about 10 mg of one or more PKC-α         inhibitors according to the present invention; and     -   b) one or more excipient.

The term “effective amount” as used herein means “an amount of one or more PKC-α inhibitors, effective at dosages and for periods of time necessary to achieve the desired result.” An effective amount may vary according to factors known in the art, such as the disease state, age, sex, and weight of the human or animal being treated. Although particular dosage regimes may be described in examples herein, a person skilled in the art would appreciated that the dosage regime may be altered to provide optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. In addition, the compositions of the present invention can be administered as frequently as necessary to achieve a therapeutic amount.

Method of Use

The present invention also relates to a method for improving cardiac contraction/relaxation parameters in heart failure patients and/or attenuating adverse cardiac remodeling and prevent or slow the progression of worsening heart failure. The present method comprises the step of administering to a human or higher mammal an effective amount of a composition comprising one or more of the PKC-α inhibitors according to the present invention.

A present invention also relates to a method of treating or preventing chronic or acute heart failure, said method comprised of the step of administering to a patient in need thereof a pharmaceutically acceptable amount of a compound according to claim 1 or a therapeutically acceptable salt thereof in combination with a drug selected that acts on the renin-angiotensin-aldosterone system, a diuretic, digoxin or a β-adrenergic receptor blockers, natriuretic peptides and inotropic agents.

The present invention also relates to the use of the 2-arylamino-4-(amino-alkylene)amino-pyrimidines or salts thereof, according to the present invention in the manufacture of a medicament for the treatment of heart disease wherein inhibition of PKC-α provides a benefit.

Procedures Assessment of PKC-α Inhibitory Activity

Measurement of PKC-α enzyme activity is performed using full-length human PKC-α enzyme (Upstate Biotechnology) at a final concentration of 0.12 μg/ml in a kinase assay buffer (0.09 mg/ml bovine serum albumin (BSA), 210 μM ethylenediaminetetraacetic acid (EDTA), 360 μM CaCl₂, 1 mM Tris-HCl, pH=7.5, 0.5 mM MgCl₂, 0.015 mg/ml phosphatidylserine and 0.015 mg/ml diacylglycerol). The reaction is initiated by addition of adenosine triphosphate (ATP; final concentration 45 μM) and a peptide substrate consisting of amino acids 28-43 (Ala-Ala-Lys-Ile-Gln-Ala-Ser-Phe-Arg-Gly-His-Met-Ala-Arg-Lys-Lys) of neurogranin (Promega; final concentration 22 μM). After a 30 minute incubation at 24° C. the reaction is terminated by adding 5 μL of the reaction mixture into 50 μL of MALDI matrix solution (5 mg/ml α-cyano-4-hydroxycinnamic acid in 50% Acetonitrile/H₂O, 0.1% TFA, 5 mM ammonium phosphate). Two microliters of the stopped reaction mixture is transferred onto a MALDI-TOF mass spectrometer target plate.

All spectra are collected on an Applied Biosystems 4700 Proteomics Analyzer MALDI-TOF MS equipped with a Nd:YAG laser (355 nm, 3 ns pulse width, 200 Hz repetition rate) in negative ion reflector mode. The system is operated with 4700 Explorer software, version 3.0. Automated acquisition parameters are adjusted to capture and average only those individual spectra within defined success criteria. Specifically, signal intensities for the substrate peptide are set to a minimum threshold of 3000 counts and a maximum intensity of 65,000 counts. This ensured that neither null spectra nor saturated spectra are averaged into the final readout. Between 1000 and 1500 laser shots are averaged for each sample. Data are collected in triplicate from 3 successive days to capture the maximum variability related to preparation of enzyme reaction, transfer of samples to MALDI target plates, data collection, and data extraction.

The isotope cluster areas for each peptide substrate and product peaks are extracted into a Microsoft Excel worksheet from the 10×10 array of spectral data simultaneously using the automated analysis function provided within the 4700 Explore software. The isotope cluster area is defined by the software algorithm based on the molecular weight and the general elemental composition of the peptides. The percent conversion (% C) of substrate to product is calculated as the cluster area of the product (P) divided by the sum of the cluster areas of the substrate (S) and the product multiplied by 100 as represented by the following equation:

${\% \mspace{14mu} C} = {\frac{P}{P + S} \times 100.}$

For dose-dependent inhibition studies, the inhibition is plotted as a % Maximal Activity (% MA). Equation one is a measure of the ratio of product to substrate then solving for the % C. However, to measure inhibition of the enzyme activity, one must measure the degree to which that activity (% C) is curtailed. Thus the dose-dependant inhibition data is plotted as % MA where the maximal activity is the % C measured in control reactions with no inhibitor as represented by the following equation:

${\% \mspace{14mu} {MA}} = {\frac{{\% \mspace{14mu} C\mspace{14mu} {with}\mspace{14mu} {inhibitor}}\mspace{14mu}}{\% \mspace{14mu} C\mspace{14mu} {with}\mspace{14mu} {no}\mspace{14mu} {inhibitor}} \times 100.}$

Evaluation of PKCα Inhibitors in Cardiomyocytes

Determination of PKCα activity in cells is determined using murine HL-1 atrial cardiac muscle cells. On day 1, HL-1 cells are plated at 18,000 cells/well in a 96-well tissue culture plate. Cells are cultured in 0.1 ml Claycomb growth medium (without norepinephrine) supplemented with 10% fetal bovine serum, 200 mM glutamine and 1% antibiotic/antimycotic. On day 2, cells are washed 1× with 100 μl of phosphate buffered saline (PBS) and the medium is replaced with 100 μl serum free Claycomb medium supplemented with 200 mM glutamine. For compound testing, the medium is removed and replaced with serum free Claycomb medium supplemented with 200 mM glutamine containing different concentrations of compound at a final volume of 50 μl. Compounds are dissolved in 100% dimethylsulfoxide (DMSO) and final DMSO concentrations are maintained at 0.5%. Plates are then incubated for 30 minutes at 37° C. in a 5% CO₂ incubator. The medium is then removed and the plates are rinsed 1× with ice-cold 100 μl PBS. The PBS is removed and replaced with 10 μl of ice-cold lysis buffer consisting of B-PERII detergent (Pierce) diluted 1:1 in distilled water and including a final concentration of 0.3% β-mercaptoethanol, 50 μg/ml phenylmethylsulfonylfluoride (PMSF) 10 mM benzamidine, 10 nM okadaic acid, 20 μg/ml leupeptin and 20 μg/ml soybean trypsin inhibitor. The plates are gently mixed for 10-20 minutes at 4° C. Next, 90 μl of coactivation buffer, consisting of 0.1 mg/ml BSA, 250 μM EDTA, 400 μM CaCl₂, is added to each well. Twenty-five microliters of the cell lysate/coactivation buffer solution is removed from each well and the enzyme activity measured by addition of 25 μl of substrate solution, consisting of 0.1 mg/ml bovine serum albumin, 235 μM EDTA, 400 μM CaCl₂, 1 mM Tris-HCl, pH=7.5, 0.5 mM MgCl₂, 0.015 mg/ml phosphatidylserine and 0.015 mg/ml diacylglycerol, 20 μM ATP and 2 μM of an octapeptide fragment of the EGF receptor (Arg-Lys-Arg-Thr-Leu-Arg-Arg-Leu). After a 30 minute incubation at 24° C. the reaction is terminated by adding 5 μL of the reaction mixture into 50 μL of MALDI matrix solution (5 mg/ml α-cyano-4-hydroxycinnamic acid in 50% Acetonitrile/H₂O, 0.1% TFA, 5 mM ammonium phosphate). Two microliters of the stopped reaction mixture is transferred onto a MALDI-TOF mass spectrometer target plate. Dose-dependent inhibition is measured by mass spectrometry as % maximal activity as described above for the isolated PKCα inhibition assays.

In Vivo Evaluation of PKCα Inhibitors in the Anesthetized Rat

Selected PKCα inhibitors are evaluated in rats with acute heart failure (HF) after myocardial infarction (MI) for effects on cardiac contractility and hemodynamics. Male, Sprague-Dawley rats are anesthetized with isoflurane, intubated, placed on ventilators and maintained at a surgical plane of anesthesia during the course of the experiment. The animals are instrumented, for the measurement of left ventricular function (+dP/dt, LVDP), arterial blood pressure, and the ECG is monitored for the incidence of arrhythmias. A thoracotomy is performed at the fourth intercostal space to visualize the heart, the pericardium is opened and a suture is placed around the left anterior descending (LAD) coronary artery approximately 3-4 mm from its origin. When hemodynamic values are stabilized, the LAD is permanently ligated to induce a myocardial infarction. Severe arrhythmia are treated with the administration of lidocaine. Typically, cardiac function stabilized approximately 40-60 min after ligation and baseline hemodynamic values are measured. N²-(3-chlorophenyl)-N⁴-(3-(dimethylamino)propyl)pyrimidine-2,4-diamine, 100 and 300 nmol/kg/min for 10 min each dose, and hemodynamic parameters are measured after each dose. The effects of treatment are normalized to pre-treatment baseline values and expressed as a percentage. Statistical significance (p<0.05) is evaluated using one-way ANOVA and Dunnett's multiple comparison test.

In Vivo Evaluation of PKCα Inhibitors in the Anesthetized Rat

Selected PKCα inhibitors are evaluated in rats with myocardial infarction (MI) for effects on cardiac contractility and hemodynamics.

Male, Sprague-Dawley or Lewis rats weighing between 225-500 gm are anesthetized with isoflurane and an MI is induced as follows. A thoracotomy is performed at the fourth intercostal space to visualize the heart, the pericardium is opened and a suture is placed around the left anterior descending (LAD) coronary artery approximately 3-4 mm from its origin. When hemodynamic values are stabilized, the LAD is permanently ligated to induce a myocardial infarction. Severe arrhythmias are treated with the administration of lidocaine. Typically, cardiac function stabilized approximately 40-60 min after ligation and baseline hemodynamic values are measured.

The effects of inhibitors on cardiac contractility and hemodynamics are evaluated in MI rats as follows. The animals are anesthetized with isoflurane. A femoral artery is isolated and cannulated for the measurement of systemic blood pressure. A jugular vein is isolated and cannulated for the intravenous infusion of inhibitor. The right carotid artery is isolated and a Millar conductance catheter is inserted to the left ventricle (LV) of the heart. The LV systolic pressure, end-diastolic pressure, +dP/dt_(max), −dP/dt_(min), and heart rate are derived from the LV pressure waveform. Mean arterial blood pressure is derived from the systemic blood pressure waveform. Data are recorded continuously and derived using computerized data acquisition software (Notocord or Powerlab). After a period of stabilization, PKC-α inhibitors are infused at the following infusion doses in MI rats: 10, 30, 100, 300 and 1000 nmol/kg/min. The infusion of each dose is allowed to run for at least five minutes. At the end of the test infusions, 5.0 μg/kg/min of dobutamine is infused. The effects of treatment are normalized to pretreatment baseline values and expressed as a percentage. Statistical significance (p<0.05) is evaluated using a one-way ANOVA and Dunnett's multiple comparison test.

Table IV provides non-limiting examples of PKC-α IC₅₀ values for representative compounds of the present invention.

TABLE IV PKC-α IC₅₀ No. Compound (nM) 1 N²-(3-chlorophenyl)-N⁴-(3-aminopropyl)pyrimidine-2,4-diamine 312 2 N²-(3-chlorophenyl)-N⁴-[3-(methylamino)propyl]pyrimidine-2,4- 4 diamine 3 N²-(3-chlorophenyl)-N⁴-[3-(dimethylamino)propyl]pyrimidine-2,4- 0.8 diamine 6 N²-(3-chlorophenyl)-N⁴-[3-(diethylamino)propyl]pyrimidine-2,4- 1 diamine 15 N²-(3,4-dichlorophenyl)-N⁴-[3-(dimethylamino)propyl]pyrimidine- 15 2,4-diamine 21 N²-(3-chloro-4-methylphenyl)-N⁴-[3-(dimethylamino)propyl]- 3 pyrimidine-2,4-diamine 25 N²-(3-chloro-2-fluorophenyl)-N⁴-[3-(dimethylamino)propyl]- 23 pyrimidine-2,4-diamine 27 N²-(3-chloro-4-fluorophenyl)-N⁴-[3-(dimethylamino)propyl]- 8 pyrimidine-2,4-diamine 33 N²-(3,4-difluorophenyl)-N⁴-[3-(dimethylamino)propyl]-pyrimidine- 65 2,4-diamine 39 N²-(3-trifluoromethylphenyl)-N⁴-[3-(dimethylamino)propyl]- 0.5 pyrimidine-2,4-diamine 45 N²-(3-trifluoromethyl-4-chlorophenyl)-N⁴-[3-(dimethylamino)- 3 propyl]pyrimidine-2,4-diamine 51 N²-(3-methoxyphenyl)-N⁴-[3-(dimethylamino)propyl]pyrimidine-2,4- 14 diamine 55 N²-(3-methylphenyl)-N⁴-[3-(dimethylamino)propyl]pyrimidine-2,4- 40 diamine 58 N²-(3-methylphenyl)-N⁴-[3-(dimethylamino)propyl]pyrimidine-2,4- 93 diamine 69 N²-(3-chlorophenyl)-N⁴-[2-(dimethylamino)ethyl]pyrimidine-2,4- 83 diamine 81 N²-(3-chlorophenyl)-N⁴-[4-(dimethylamino)butyl]pyrimidine-2,4- 12 diamine 87 N²-(biphenyl-3-yl)-N⁴-[3-(dimethylamino)propyl]pyrimidine-2,4- 0.4 diamine 105 N²-[3-(2-methyl-thiazol-4-yl)phenyl]-N⁴-[3-(dimethylamino)- 48 propyl]pyrimidine-2,4-diamine

While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention. 

1. A compound having the formula:

wherein R is a unit having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄ substituted or unsubstituted linear, branched, or cyclic alkyl; L is a linking unit having the formula: —[C(R^(4a)R^(4b))]_(n)— each R^(4a) and R^(4b) is independently chosen from: i) hydrogen; or ii) C₁-C₄ linear, branched, or cyclic alkyl; the index n is from 1 to 4; and R¹ is substituted or unsubstituted aryl; excluding the compounds N²-(4-chlorophenyl)-N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamine, N²-(4-chlorophenyl)-N⁴-[(3-diethylamino)propyl]-pyrimidine-2,4-diamine, and N²-(4-methylphenyl)-N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamine.
 2. A compound according to claim 1 wherein R² and R³ are each independently chosen from hydrogen, methyl, ethyl, n-propyl, iso-propyl, cyclopropyl, n-butyl, iso-butyl, sec-butyl, and tert-butyl.
 3. A compound according to claim 1 wherein R is a unit chosen from —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃), —N(CH₂CH₃)₂, —N(CH₃)(CH₂CH₃), —NH(CH₂CH₂CH₃), —N(CH₃)(CH₂CH₂CH₃), —N(CH₂CH₃)(CH₂CH₂CH₃), and —N(CH₂CH₂CH₃)₂.
 4. A compound according to claim 3 wherein R¹ is a unit chosen from —NH₂, —NHCH₃, —N(CH₃)₂, —NH(CH₂CH₃), and —N(CH₂CH₃)₂.
 5. A compound according to claim 1 wherein R¹ is a phenyl unit substituted with one or more units chosen from: i) C₁-C₄ linear, branched, or cyclic alkyl; ii) halogen; iii)-OR⁶; iv)-SO₂N(R⁶)₂; v) —CH_(m)X_(n); wherein each X is independently F, Cl, Br, or I, m is from 0 to 2, m+n=3; and vi) —NO₂; each R⁶ is independently hydrogen, C₁-C₄ linear, branched, or cyclic alkyl, or phenyl.
 6. A compound according to claim 5 wherein R¹ is chosen from 3-chlorophenyl, 4-chlorophenyl, 3,4-dichlorophenyl, 3-chloro-4-methylphenyl, 3-chloro-4-fluorophenyl, 3,4-difluorophenyl, 3-trifluoromethylphenyl, 3-trifluoromethyl-4-chlorophenyl, 3-methoxyphenyl, 3-methylphenyl, 3-ethylphenyl, and 3-isopropylphenyl.
 7. A compound according to claim 5 wherein R¹ is chosen from 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4-trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6-trifluorophenyl, 2-chlorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, 2,6-dichlorophenyl, 2,3,4-trichlorophenyl, 2,3,5-trichlorophenyl, 2,3,6-trichlorophenyl, 2,4,5-trichlorophenyl, and 2,4,6-trichlorophenyl.
 8. A compound according to claim 5 wherein R¹ is chosen from 2-methylphenyl, 4-methylphenyl, 2,3-dimethylphenyl, 2,4-dimethylphenyl, 2,5-dimethyl-phenyl, 2,6-dimethylphenyl, 3,4-dimethyl-phenyl, 2,3,4-trimethylphenyl, 2,3,5-trimethyl-phenyl, 2,3,6-trimethylphenyl, 2,4,5-trimethylphenyl, 2,4,6-trimethylphenyl, 2-ethylphenyl, 4-ethylphenyl, 2,3-diethylphenyl, 2,4-diethylphenyl, 2,5-diethylphenyl, 2,6-diethylphenyl, 3,4-diethylphenyl, 2,3,4-triethylphenyl, 2,3,5-triethylphenyl, 2,3,6-triethylphenyl, 2,4,5-triethylphenyl, and 2,4,6-triethylphenyl.
 9. A compound according to claim 5 wherein R¹ is chosen from 2-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 2,4-dimethoxyphenyl, 2,5-dimethoxyphenyl, 2,6-dimethoxyphenyl, 3,4-dimethoxyphenyl, 2,3,4-trimethoxyphenyl, 2,3,5-trimethoxyphenyl, 2,3,6-trimethoxy-phenyl, 2,4,5-trimethoxyphenyl, 2,4,6-trimethoxyphenyl, 2-hydroxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 2,3-dihydroxyphenyl, 2,4-dihydroxyphenyl, 2,5-dihydroxyphenyl, 2,6-dihydroxyphenyl, 3,4-dihydroxy-phenyl, 2,3,4-trihydroxyphenyl, 2,3,5-trihydroxy-phenyl, 2,3,6-trihydroxyphenyl, 2,4,5-trihydroxyphenyl, and 2,4,6-trihydroxyphenyl.
 10. A compound having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄ linear or branched alkyl; and R⁵ is chosen from: i) C₁-C₄ linear, branched, or cyclic alkyl; ii) halogen; iii)-OR⁶; iv) —SO₂N(R⁶)₂; v) —CH_(m)X_(n); wherein each X is independently F, Cl, Br, or I, m is from 0 to 2, m+n=3; vi) —NO₂; and vii) phenyl; each R⁶ is independently hydrogen, or C₁-C₄ linear, branched, or cyclic alkyl; excluding N²-(4-chlorophenyl)-N⁴-[(3-diethylamino)propyl]-pyrimidine-2,4-diamine.
 11. A compound according to claim 10 wherein R² and R³ are each independently hydrogen, methyl, and ethyl.
 12. A compound according to claim 10 wherein R⁵ is chosen from 3-chloro, 4-chloro, 3,4-dichloro, 3-chloro-4-methyl-, 3-chloro-4-fluoro, 3,4-difluoro, 3-trifluoro-methyl, 3-trifluoromethyl-4-chloro, 3-methoxy, 3-methyl, 3-ethyl, and 3-iso-propyl.
 13. A compound according to claim 10 wherein R⁵ is chosen from 2-fluoro, 3-fluoro, 4-fluoro, 2,3-difluoro, 2,4-difluoro, 2,5-difluoro, 2,6-difluoro, 2-chloro, 2,3-dichloro, 2,4-dichloro, 2,5-dichloro and, 2,6-dichloro.
 14. A compound according to claim 10 wherein R⁵ is chosen from 2-methoxy, 4-methoxy, 2,3-dimethoxy, 2,4-dimethoxy, 2,5-dimethoxy, 2,6-dimethoxy, 3,4-dimethoxy, 2-hydroxy, 3-hydroxy, 4-hydroxy, 2,3-dihydroxy, 2,4-dihydroxy, 2,5-dihydroxy, 2,6-dihydroxy, and 3,4-dihydroxy.
 15. A compound having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄ linear or branched alkyl; and R⁵ is chosen from: i) C₁-C₄ linear, branched, or cyclic alkyl; ii) halogen; iii) —OR⁶; iv) —SO₂N(R⁶)₂; v) —CH_(m)X_(n); wherein each X is independently F, Cl, Br, or I, m is from 0 to 2, m+n=3; vi) —NO₂; and vii) phenyl; each R⁶ is independently hydrogen, or C₁-C₄ linear, branched, or cyclic alkyl; and L is chosen from: i) —CH₂—, methylene; ii) —CH₂CH₂—, ethylene; iii) —CH(CH₃)CH₂—, 1-propylene; iv) —CH₂CH(CH₃)—, 2-propylene; v) —CH₂CH₂CH₂CH₂—, butylene. vi) —CH(CH₃)CH₂CH₂—, 1-butylene; vii) —CH₂CH(CH₃)CH₂—, 2-butylene; and viii) —CH₂C(CH₃)₂CH₂—, 2,2-dimethylpropylene; excluding N²-(4-chlorophenyl)-N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamine and N²-(4-methylphenyl)-N⁴-[(2-diethylamino)ethyl]-pyrimidine-2,4-diamine.
 16. A composition comprising: A) a compound having the formula:

wherein R is a unit having the formula:

R² and R³ are each independently chosen from: i) hydrogen; or ii) C₁-C₄ linear, branched, or cyclic alkyl; L is a linking unit having the formula: —[C(R^(4a)R^(4b))]_(n)— each R^(4a) and R^(4b) is independently chosen from: i) hydrogen; or ii) C₁-C₄ linear, branched, or cyclic alkyl; the index n is from 1 to 4; and R¹ is aryl; and B) the balance carriers and excipients.
 17. A method for treating or preventing a disease or medical condition selected from diabetes, numerous forms of cancer, microalbinuria, endothelial dysfunction, cerebrovascular disease, stroke, coronary heart disease, cardiovascular disease and sequela (e.g. arrhythmia, sudden death, increased infarct size, congestive heart failure, angina), myocardial ischemic states, hypertension, lipid disorders, ischemia-reperfusion injury, atherosclerosis, peripheral artery/vascular disease, microvascular complications of diabetes (neuropathy, nephropathy, retinopathy), restenosis, renal disease, blood coagulation disorders, inflammatory diseases, cardiac hypertrophy, dilated cardiomyopathy, ischemic injury and suboptimal mitogen stimulation said method comprised of the steps of administering to a patient in need thereof a therapeutic amount of a compound according to claim
 1. 18. A method for treating acute heart failure comprising administering to a human an effective amount of a composition comprising one or more of the PKC-α inhibitors according to claim
 1. 19. A method for chronic heart failure comprising administering to a human an effective amount of a composition comprising one or more of the PKC-α inhibitors according to claim
 1. 20. A method of treating or preventing chronic or acute heart failure, said method comprised of the step of administering to a patient in need thereof a pharmaceutically acceptable amount of a compound according to claim 1 or a therapeutically acceptable salt thereof in combination with a drug selected that acts on the renin-angiotensin-aldosterone system, a diuretic, digoxin or a β-adrenergic receptor blockers, natriuretic peptides and inotropic agents.
 21. The method of claim 20 wherein the drug used in combination with a compound according to claim 1 is a renin-angiotensin-aldosterone system selected from an ACEI, ARB, or aldosterone inhibitor. 